REPLICATION VS POLYMERASE CHAIN REACTION
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PCR allows for targeted amplification of specific DNA fragments. It involves repeated cycles of DNA denaturation by heating and cooling, primer annealing, and extension of the DNA strand by a DNA polymerase. Key steps include separation of double stranded DNA at 95°C, annealing of primers at 40-65°C, and extension of the DNA chain by DNA polymerase at 72°C. PCR amplifies the target DNA sequence up to 100,000-fold, enabling detection of rare DNA sequences. It has largely replaced other methods for DNA analysis and molecular cloning due to its specificity, sensitivity, speed, and ability to quantitatively analyze DNA.
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Purification of dna from living cells
1. Purification of total DNA from bacterial cells involves growing and harvesting the bacterial culture, disrupting the cells, removing cellular components except DNA, and concentrating the DNA.
2. Cells are disrupted through chemical or physical lysis methods. Chemical lysis uses lysozyme and EDTA to weaken the cell wall and SDS to disrupt the cell membrane.
3. Phenol-chloroform extraction is used to remove proteins from the cell extract by partitioning them into the organic phase, leaving DNA and RNA in the aqueous phase. Ethanol precipitation is then used to concentrate the purified DNA.
DNA: structure, organization and function
Dr. Ifat Ara Begum's document provides an overview of DNA structure, organization, and function. Some key points include:
- DNA is a polymer of deoxyribonucleotides connected by phosphodiester bonds that carries the genetic instructions for life.
- The DNA double helix consists of two antiparallel strands connected by hydrogen bonds between complementary nucleotide base pairs.
- DNA is organized into chromosomes through progressive coiling and folding. Chromosomes are highly condensed and visible during cell division.
- DNA contains both coding and non-coding regions. While genes only account for about 1.5% of DNA, non-coding DNA plays important roles in packaging, expression, and regulation of genes
Polymerase chain reaction
Polymerase chain reaction (PCR) was invented in 1984 by Kary Mullis and revolutionized biological research. PCR amplifies a specific DNA sequence using heat-stable DNA polymerase and primers. The process involves denaturation of DNA, annealing of primers, and extension of primers to exponentially amplify the target sequence. Variations of PCR include real-time PCR, reverse transcription PCR, asymmetric PCR, and site-directed mutagenesis PCR, which are used for applications such as detecting pathogens and inducing mutations.
Molecular biology
Molecular biology is a branch of science concerning biological activity at the molecular level.
The field of molecular biology overlaps with biology and chemistry and in particular, genetics and biochemistry.
A key area of molecular biology concerns understanding how various cellular systems interact in terms of the way DNA, RNA and protein synthesis function.
Molecular biology is the study of molecular underpinnings of the process of replication, transcription and translation of the genetic material.
Random Amplified Polymorphic Dna
This document summarizes the random amplified polymorphic DNA (RAPD) technique. RAPD is a type of PCR that uses random nucleotide primers to amplify unknown regions of genomic DNA. It was developed in 1991 and involves using single, short random primers to amplify random DNA segments. The amplified products are then separated via gel electrophoresis and visualized. RAPD is a quick and inexpensive molecular marker technique that requires no prior DNA sequence knowledge, but lacks reproducibility and produces dominant markers. Its applications include assessing genetic diversity, mapping genomes, and use in breeding and evolutionary studies.
Polymerase Chain Reaction(PCR) Likhith K
The advent of the polymerase chain reaction (PCR) radically transformed biological science from the time it was first discovered (Mullis, 1990). For the first time, it allowed for specific detection and production of large amounts of DNA. PCR-based strategies have propelled huge scientific endeavors such as the Human Genome Project. The technique is currently widely used by clinicians and researchers to diagnose diseases, clone and sequence genes, and carry out sophisticated quantitative and genomic studies in a rapid and very sensitive manner. One of the most important medical applications of the classical PCR method is the detection of pathogens. In addition, the PCR assay is used in forensic medicine to identify criminals. Because of its widespread use, it is important to understand the basic principles of PCR and how its use can be modified to provide for sophisticated analysis of genes and the genome
Random amplified polymorphic DNA-RAPD.pptx
Random Amplified Polymorphic DNA is a type of PCR in which the segments of DNA that are amplified are random.
Williams et al. (1990) developed this technique using very short primers to generate random fragments from template DNAs.
RAPD fragments can be separated and used as genetic markers or a kind of DNA fingerprinting.
Principle
The standard RAPD technology utilizes short synthetic oligonucleotides (10 bases long) of random sequences as primers to amplify nanogram amounts of total genomic DNA under low annealing temperatures by PCR. Amplification products are generally separated on agarose gels and stained with ethidium bromide.
At an appropriate annealing temperature during the thermal cycle, oligonucleotide primers of random sequence bind several priming sites on the complementary sequences in the template genomic DNA and produce discrete DNA products if these priming sites are within an amplifiable distance of each other.
The profile of amplified DNA primarily depends on nucleotide sequence homology between the template DNA and oligonucleotide primer at the end of each amplified product. Nucleotide variation between different sets of template DNAs will result in the presence or absence of bands because of changes in the priming sites.
Procedure
1. DNA is made single stranded by raising the temperature to 940C.
2. In second step, temperature is lowered to 40- 650C which results in annealing of the primer to their target sequences on the template DNA.
3. Temperature is chosen where the activity of the thermostable Taq DNA polymerase is optimal.
4. The polymerase now extends the 3’ ends of the DNA primer hybrids towards the outer primer binding site.
5. Repeating these three step cycles 40 to 50 times results in the exponential amplification of the target between the 5’ ends of the two primer binding sites.
6. Amplification products are separated by gel electrophoresis and visualized by ethidium bromide staining.
Pyrosequencing 454
The document describes the process of 454-pyrosequencing for next generation sequencing. It involves the following main steps: 1) obtaining a sample and fragmenting its DNA library, 2) preparing the library with adapters and denaturing, 3) annealing fragments to beads in an emulsion to clonally amplify each fragment, 4) breaking the emulsion and loading beads onto a picotiter plate for sequencing, 5) performing pyrosequencing chemistry using enzymes and cameras to detect light signals from nucleotide washes to determine sequences. The data is then processed and analyzed for assembly, mapping, and variant detection.
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Purification of dna from living cells
1. Purification of total DNA from bacterial cells involves growing and harvesting the bacterial culture, disrupting the cells, removing cellular components except DNA, and concentrating the DNA.
2. Cells are disrupted through chemical or physical lysis methods. Chemical lysis uses lysozyme and EDTA to weaken the cell wall and SDS to disrupt the cell membrane.
3. Phenol-chloroform extraction is used to remove proteins from the cell extract by partitioning them into the organic phase, leaving DNA and RNA in the aqueous phase. Ethanol precipitation is then used to concentrate the purified DNA.
DNA: structure, organization and function
Dr. Ifat Ara Begum's document provides an overview of DNA structure, organization, and function. Some key points include:
- DNA is a polymer of deoxyribonucleotides connected by phosphodiester bonds that carries the genetic instructions for life.
- The DNA double helix consists of two antiparallel strands connected by hydrogen bonds between complementary nucleotide base pairs.
- DNA is organized into chromosomes through progressive coiling and folding. Chromosomes are highly condensed and visible during cell division.
- DNA contains both coding and non-coding regions. While genes only account for about 1.5% of DNA, non-coding DNA plays important roles in packaging, expression, and regulation of genes
Polymerase chain reaction
Polymerase chain reaction (PCR) was invented in 1984 by Kary Mullis and revolutionized biological research. PCR amplifies a specific DNA sequence using heat-stable DNA polymerase and primers. The process involves denaturation of DNA, annealing of primers, and extension of primers to exponentially amplify the target sequence. Variations of PCR include real-time PCR, reverse transcription PCR, asymmetric PCR, and site-directed mutagenesis PCR, which are used for applications such as detecting pathogens and inducing mutations.
Molecular biology
Molecular biology is a branch of science concerning biological activity at the molecular level.
The field of molecular biology overlaps with biology and chemistry and in particular, genetics and biochemistry.
A key area of molecular biology concerns understanding how various cellular systems interact in terms of the way DNA, RNA and protein synthesis function.
Molecular biology is the study of molecular underpinnings of the process of replication, transcription and translation of the genetic material.
Random Amplified Polymorphic Dna
This document summarizes the random amplified polymorphic DNA (RAPD) technique. RAPD is a type of PCR that uses random nucleotide primers to amplify unknown regions of genomic DNA. It was developed in 1991 and involves using single, short random primers to amplify random DNA segments. The amplified products are then separated via gel electrophoresis and visualized. RAPD is a quick and inexpensive molecular marker technique that requires no prior DNA sequence knowledge, but lacks reproducibility and produces dominant markers. Its applications include assessing genetic diversity, mapping genomes, and use in breeding and evolutionary studies.
Polymerase Chain Reaction(PCR) Likhith K
The advent of the polymerase chain reaction (PCR) radically transformed biological science from the time it was first discovered (Mullis, 1990). For the first time, it allowed for specific detection and production of large amounts of DNA. PCR-based strategies have propelled huge scientific endeavors such as the Human Genome Project. The technique is currently widely used by clinicians and researchers to diagnose diseases, clone and sequence genes, and carry out sophisticated quantitative and genomic studies in a rapid and very sensitive manner. One of the most important medical applications of the classical PCR method is the detection of pathogens. In addition, the PCR assay is used in forensic medicine to identify criminals. Because of its widespread use, it is important to understand the basic principles of PCR and how its use can be modified to provide for sophisticated analysis of genes and the genome
Random amplified polymorphic DNA-RAPD.pptx
Random Amplified Polymorphic DNA is a type of PCR in which the segments of DNA that are amplified are random.
Williams et al. (1990) developed this technique using very short primers to generate random fragments from template DNAs.
RAPD fragments can be separated and used as genetic markers or a kind of DNA fingerprinting.
Principle
The standard RAPD technology utilizes short synthetic oligonucleotides (10 bases long) of random sequences as primers to amplify nanogram amounts of total genomic DNA under low annealing temperatures by PCR. Amplification products are generally separated on agarose gels and stained with ethidium bromide.
At an appropriate annealing temperature during the thermal cycle, oligonucleotide primers of random sequence bind several priming sites on the complementary sequences in the template genomic DNA and produce discrete DNA products if these priming sites are within an amplifiable distance of each other.
The profile of amplified DNA primarily depends on nucleotide sequence homology between the template DNA and oligonucleotide primer at the end of each amplified product. Nucleotide variation between different sets of template DNAs will result in the presence or absence of bands because of changes in the priming sites.
Procedure
1. DNA is made single stranded by raising the temperature to 940C.
2. In second step, temperature is lowered to 40- 650C which results in annealing of the primer to their target sequences on the template DNA.
3. Temperature is chosen where the activity of the thermostable Taq DNA polymerase is optimal.
4. The polymerase now extends the 3’ ends of the DNA primer hybrids towards the outer primer binding site.
5. Repeating these three step cycles 40 to 50 times results in the exponential amplification of the target between the 5’ ends of the two primer binding sites.
6. Amplification products are separated by gel electrophoresis and visualized by ethidium bromide staining.
Pyrosequencing 454
The document describes the process of 454-pyrosequencing for next generation sequencing. It involves the following main steps: 1) obtaining a sample and fragmenting its DNA library, 2) preparing the library with adapters and denaturing, 3) annealing fragments to beads in an emulsion to clonally amplify each fragment, 4) breaking the emulsion and loading beads onto a picotiter plate for sequencing, 5) performing pyrosequencing chemistry using enzymes and cameras to detect light signals from nucleotide washes to determine sequences. The data is then processed and analyzed for assembly, mapping, and variant detection.
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The document provides information on the basics of molecular biology. It begins with a table comparing key attributes of eukaryotes and prokaryotes. It then defines molecular biology as the study of the molecular underpinnings of processes like DNA replication, transcription, and translation. The basic components involved are described as DNA, RNA, and proteins. DNA stores genetic information. RNA and proteins are involved in building and regulating cells. The processes of DNA replication, transcription, translation, and their roles are summarized.
DNA transcription & Post Transcriptional Modification
DNA is transcribed into RNA through the process of transcription. In eukaryotes, transcription is initiated when RNA polymerase binds to a promoter region near a gene. It then elongates the RNA molecule using the DNA as a template. Transcription ends when the polymerase reaches a termination sequence. The primary RNA transcript often undergoes processing like splicing, capping, polyadenylation, and editing to become a functional mRNA, tRNA, or rRNA molecule. These post-transcriptional modifications are required for gene expression.
DNA manipulation
DNA manipulating enzymes include nucleases, ligases, polymerases and modifying enzymes. Restriction endonucleases are enzymes that cut DNA molecules at specific recognition sequences. They produce either blunt or sticky ends. Restriction endonucleases are classified as type I, II or III, with type II having widespread use in recombinant DNA technology by allowing precise cutting of DNA molecules at specific sites.
PCR PPT
PCR (polymerase chain reaction) was invented in 1987 by Kary Mullis to specifically amplify a single DNA sequence. It takes advantage of DNA replication by using DNA polymerase, primers, and thermocycling to exponentially amplify the target sequence. PCR is now widely used in applications such as cloning, forensics, disease detection, and more due to its ability to amplify small amounts of DNA.
Chromosome structure
1. Viral genomes contain DNA or RNA and are packaged into capsids through assembly processes. Bacterial chromosomes contain genes and other sequences compacted by looping and supercoiling.
2. Eukaryotic chromosomes vary greatly in size and contain genes and other sequences. Their DNA must be highly compacted to fit in the nucleus.
3. Eukaryotic DNA wraps around histone proteins to form nucleosomes, which further compact to form chromatin fibers and loop domains anchored to the nuclear matrix. Additional compaction occurs during cell division through condensin and cohesin proteins.
Mitochondrial dna
Mitochondrial DNA is circular DNA located in the mitochondria of cells. It is 16kb in size and encodes 37 genes. Mitochondrial DNA is only inherited from the mother and differs from nuclear DNA in several key ways, such as being maternally inherited only. Mutations in mitochondrial DNA can cause diseases often involving the central nervous system or musculoskeletal system. The high mutation rate of mitochondrial DNA is due to lack of protective histones and DNA repair mechanisms.
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...
This slidedeck introduces the concepts of real-time PCR and how to conduct a real-time PCR assay. The topics that are covered include an overview of real-time PCR chemistries, protocols, quantification methods, real-time PCR applications and factors for success.
Dna quantification
this section helps students how to quanify the isolated DNA by spectrophotometer. specially life life science fields such as biotechnology, biology, and medical laboratory
Structure Of Dna 1201627173171032 3
DNA is a long, double-stranded molecule made up of nucleotides that functions to store and retrieve genetic information. It consists of a sugar, phosphate, and one of four nitrogenous bases. The bases on each strand form complementary base pairs with the other strand through hydrogen bonding between adenine and thymine or cytosine and guanine. This base pairing allows DNA to store and replicate genetic information that is used to direct the synthesis of proteins and transfer hereditary traits.
Dna sequencing
DNA sequencing is the process of determining the order of nucleotides in DNA. There are several methods of DNA sequencing including conventional, cycle sequencing, automated sequencing, and pyrosequencing. Conventional methods include chemical degradation and chain termination. Chemical degradation uses base-specific chemical reactions to cleave DNA fragments for sequencing. Chain termination uses DNA polymerase and dideoxynucleotides to terminate DNA strand extension for sequencing. Cycle sequencing applies the chain termination method to PCR for linear amplification of sequencing products. Automated sequencing uses fluorescence labeling for high-throughput sequencing. Pyrosequencing sequences DNA by detecting pyrophosphate release during polymerase nucleotide incorporation without electrophoresis.
RNA structure
Medical Biochemistry , MBBS, BDS and General Biochemistry, Types of RNA, mRNA, tRNA, rRNA, SnRNA, HnRNA, SiRNA, MiRNA, ScRNA
Translation
1) Translation is the process by which the genetic code stored in mRNA is used to direct the synthesis of proteins from amino acids. It involves decoding the mRNA codon triplet into the corresponding amino acid.
2) The key components of translation are tRNAs, ribosomes, mRNA, amino acids, and various protein factors. tRNAs carry amino acids and recognize mRNA codons through base pairing of its anticodon.
3) Translation occurs through three main steps - initiation, elongation, and termination. In initiation, the small ribosomal subunit binds mRNA and a start codon. In elongation, amino acids are added one by one. In termination, release factors recognize stop codons and cause dissociation of the ribosome and
Structure of dna
DNA is composed of nucleotides that contain nitrogen bases, sugars, and phosphates. The order of these nucleotides determines the genetic code. DNA exists as a double helix structure with two complementary strands joined by hydrogen bonds between nitrogen bases on each strand. This double helix structure allows DNA to efficiently store and replicate genetic information.
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History of DNA. introduction of DNA with short history and findings. different types of DNA with structures variations. A -DNA, B- DNA, C- DNA E- DNA D- DNA And Z DNA Detail information of these DNA with their comparison tables, different types of unusual DNA and sequences. Functions of DNA with their explanations . Nucleic acid chemical basis : Denaturation and annealing of DNA with factors for that. New DNA.
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DNA sequencing is a laboratory technique used to determine the exact sequence of bases (A, C, G, and T) in a DNA molecule. The DNA base sequence carries the information a cell needs to assemble protein and RNA molecules. DNA sequence information is important to scientists investigating the functions of genes.
In medicine, DNA sequencing is used for a range of purposes, including diagnosis and treatment of diseases. In general, sequencing allows health care practitioners to determine if a gene or the region that regulates a gene contains changes, called variants or mutations, that are linked to a disorder.
Genetic Material
The genetic material of a cell or an organism refers to those materials found in the nucleus, mitochondria and cytoplasm, which play a fundamental role in determining the structure and nature of cell substances, and capable of self-propagating and variation.
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This PPT shows the general information about PCR principles and gene expression analysis. It might be useful for researchers, students working in the field of molecular biology and genomics.
MODIFYING ENZYMES
This document discusses various enzymes used for genetic engineering and DNA manipulation. It describes restriction endonucleases and DNA ligase which cut and join DNA fragments. It also discusses other DNA modifying enzymes like nucleases which degrade DNA, and polymerases which synthesize DNA copies. Specific enzymes covered in detail include DNA polymerase I, T4 DNA polymerase, T7 DNA polymerase, terminal transferase, T4 DNA ligase, and T4 RNA ligase.
Molecular biology
well, dis z again another ppt on molecular biology..
I know dis kinda luks boring bt pretty informative
thanks
let me know wat you think abt dis
don't forget to comment
Asymetric -PCR
What is PCR
Basic Requirements
Types of PCR
Asymmetric PCR
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DNA Template
Primers
Taq polymerase
Deoxynucleoside
triphosphates(dNTPs)
Buffer solution
Divalent cations(eg.Mg2+ )
PCR-SlideShare
PCR (polymerase chain reaction) is an in vitro technique used to amplify a specific region of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate the DNA strands and allow primers to anneal, followed by extension of the primers by a thermostable DNA polymerase. Kary Mullis developed PCR in 1985 and was awarded the Nobel Prize for Chemistry in 1993. Requirements for PCR include a DNA template, primers, Taq polymerase enzyme, dNTPs, buffer solution and magnesium ions. There are several applications and variations of PCR including quantitative real-time PCR, reverse transcription PCR, and inverse PCR.
Presentation of plant protoplasts
Protoplasts are plant cells that have had their cell walls removed, allowing for genetic manipulation and fusion. The document details methods for isolating protoplasts from plant tissues using either mechanical or enzymatic methods. Once isolated, protoplast viability and density can be determined before attempting to culture the protoplasts and induce cell wall regeneration. Applications include somatic hybridization through protoplast fusion to transfer genes between species.
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BASICS OF MOLECULAR BIOLOGY
The document provides information on the basics of molecular biology. It begins with a table comparing key attributes of eukaryotes and prokaryotes. It then defines molecular biology as the study of the molecular underpinnings of processes like DNA replication, transcription, and translation. The basic components involved are described as DNA, RNA, and proteins. DNA stores genetic information. RNA and proteins are involved in building and regulating cells. The processes of DNA replication, transcription, translation, and their roles are summarized.
DNA transcription & Post Transcriptional Modification
DNA is transcribed into RNA through the process of transcription. In eukaryotes, transcription is initiated when RNA polymerase binds to a promoter region near a gene. It then elongates the RNA molecule using the DNA as a template. Transcription ends when the polymerase reaches a termination sequence. The primary RNA transcript often undergoes processing like splicing, capping, polyadenylation, and editing to become a functional mRNA, tRNA, or rRNA molecule. These post-transcriptional modifications are required for gene expression.
DNA manipulation
DNA manipulating enzymes include nucleases, ligases, polymerases and modifying enzymes. Restriction endonucleases are enzymes that cut DNA molecules at specific recognition sequences. They produce either blunt or sticky ends. Restriction endonucleases are classified as type I, II or III, with type II having widespread use in recombinant DNA technology by allowing precise cutting of DNA molecules at specific sites.
PCR PPT
PCR (polymerase chain reaction) was invented in 1987 by Kary Mullis to specifically amplify a single DNA sequence. It takes advantage of DNA replication by using DNA polymerase, primers, and thermocycling to exponentially amplify the target sequence. PCR is now widely used in applications such as cloning, forensics, disease detection, and more due to its ability to amplify small amounts of DNA.
Chromosome structure
1. Viral genomes contain DNA or RNA and are packaged into capsids through assembly processes. Bacterial chromosomes contain genes and other sequences compacted by looping and supercoiling.
2. Eukaryotic chromosomes vary greatly in size and contain genes and other sequences. Their DNA must be highly compacted to fit in the nucleus.
3. Eukaryotic DNA wraps around histone proteins to form nucleosomes, which further compact to form chromatin fibers and loop domains anchored to the nuclear matrix. Additional compaction occurs during cell division through condensin and cohesin proteins.
Mitochondrial dna
Mitochondrial DNA is circular DNA located in the mitochondria of cells. It is 16kb in size and encodes 37 genes. Mitochondrial DNA is only inherited from the mother and differs from nuclear DNA in several key ways, such as being maternally inherited only. Mutations in mitochondrial DNA can cause diseases often involving the central nervous system or musculoskeletal system. The high mutation rate of mitochondrial DNA is due to lack of protective histones and DNA repair mechanisms.
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...
This slidedeck introduces the concepts of real-time PCR and how to conduct a real-time PCR assay. The topics that are covered include an overview of real-time PCR chemistries, protocols, quantification methods, real-time PCR applications and factors for success.
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this section helps students how to quanify the isolated DNA by spectrophotometer. specially life life science fields such as biotechnology, biology, and medical laboratory
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DNA is a long, double-stranded molecule made up of nucleotides that functions to store and retrieve genetic information. It consists of a sugar, phosphate, and one of four nitrogenous bases. The bases on each strand form complementary base pairs with the other strand through hydrogen bonding between adenine and thymine or cytosine and guanine. This base pairing allows DNA to store and replicate genetic information that is used to direct the synthesis of proteins and transfer hereditary traits.
Dna sequencing
DNA sequencing is the process of determining the order of nucleotides in DNA. There are several methods of DNA sequencing including conventional, cycle sequencing, automated sequencing, and pyrosequencing. Conventional methods include chemical degradation and chain termination. Chemical degradation uses base-specific chemical reactions to cleave DNA fragments for sequencing. Chain termination uses DNA polymerase and dideoxynucleotides to terminate DNA strand extension for sequencing. Cycle sequencing applies the chain termination method to PCR for linear amplification of sequencing products. Automated sequencing uses fluorescence labeling for high-throughput sequencing. Pyrosequencing sequences DNA by detecting pyrophosphate release during polymerase nucleotide incorporation without electrophoresis.
RNA structure
Medical Biochemistry , MBBS, BDS and General Biochemistry, Types of RNA, mRNA, tRNA, rRNA, SnRNA, HnRNA, SiRNA, MiRNA, ScRNA
Translation
1) Translation is the process by which the genetic code stored in mRNA is used to direct the synthesis of proteins from amino acids. It involves decoding the mRNA codon triplet into the corresponding amino acid.
2) The key components of translation are tRNAs, ribosomes, mRNA, amino acids, and various protein factors. tRNAs carry amino acids and recognize mRNA codons through base pairing of its anticodon.
3) Translation occurs through three main steps - initiation, elongation, and termination. In initiation, the small ribosomal subunit binds mRNA and a start codon. In elongation, amino acids are added one by one. In termination, release factors recognize stop codons and cause dissociation of the ribosome and
Structure of dna
DNA is composed of nucleotides that contain nitrogen bases, sugars, and phosphates. The order of these nucleotides determines the genetic code. DNA exists as a double helix structure with two complementary strands joined by hydrogen bonds between nitrogen bases on each strand. This double helix structure allows DNA to efficiently store and replicate genetic information.
Deoxyribonucleic Acid (DNA)
History of DNA. introduction of DNA with short history and findings. different types of DNA with structures variations. A -DNA, B- DNA, C- DNA E- DNA D- DNA And Z DNA Detail information of these DNA with their comparison tables, different types of unusual DNA and sequences. Functions of DNA with their explanations . Nucleic acid chemical basis : Denaturation and annealing of DNA with factors for that. New DNA.
-DNA Sequencing Notes.pdf
DNA sequencing is a laboratory technique used to determine the exact sequence of bases (A, C, G, and T) in a DNA molecule. The DNA base sequence carries the information a cell needs to assemble protein and RNA molecules. DNA sequence information is important to scientists investigating the functions of genes.
In medicine, DNA sequencing is used for a range of purposes, including diagnosis and treatment of diseases. In general, sequencing allows health care practitioners to determine if a gene or the region that regulates a gene contains changes, called variants or mutations, that are linked to a disorder.
Genetic Material
The genetic material of a cell or an organism refers to those materials found in the nucleus, mitochondria and cytoplasm, which play a fundamental role in determining the structure and nature of cell substances, and capable of self-propagating and variation.
PRINCIPLES OF PCR AND GENE EXPRESSION ANALYSIS
This PPT shows the general information about PCR principles and gene expression analysis. It might be useful for researchers, students working in the field of molecular biology and genomics.
MODIFYING ENZYMES
This document discusses various enzymes used for genetic engineering and DNA manipulation. It describes restriction endonucleases and DNA ligase which cut and join DNA fragments. It also discusses other DNA modifying enzymes like nucleases which degrade DNA, and polymerases which synthesize DNA copies. Specific enzymes covered in detail include DNA polymerase I, T4 DNA polymerase, T7 DNA polymerase, terminal transferase, T4 DNA ligase, and T4 RNA ligase.
Molecular biology
well, dis z again another ppt on molecular biology..
I know dis kinda luks boring bt pretty informative
thanks
let me know wat you think abt dis
don't forget to comment
Asymetric -PCR
What is PCR
Basic Requirements
Types of PCR
Asymmetric PCR
Applications of PCR
Advantages of PCR
Limitations of PCR
DNA Template
Primers
Taq polymerase
Deoxynucleoside
triphosphates(dNTPs)
Buffer solution
Divalent cations(eg.Mg2+ )
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DNA transcription & Post Transcriptional Modification
DNA transcription & Post Transcriptional Modification
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...
Introduction to Real Time PCR (Q-PCR/qPCR/qrt-PCR): qPCR Technology Webinar S...
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PCR-SlideShare
PCR (polymerase chain reaction) is an in vitro technique used to amplify a specific region of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate the DNA strands and allow primers to anneal, followed by extension of the primers by a thermostable DNA polymerase. Kary Mullis developed PCR in 1985 and was awarded the Nobel Prize for Chemistry in 1993. Requirements for PCR include a DNA template, primers, Taq polymerase enzyme, dNTPs, buffer solution and magnesium ions. There are several applications and variations of PCR including quantitative real-time PCR, reverse transcription PCR, and inverse PCR.
Presentation of plant protoplasts
Protoplasts are plant cells that have had their cell walls removed, allowing for genetic manipulation and fusion. The document details methods for isolating protoplasts from plant tissues using either mechanical or enzymatic methods. Once isolated, protoplast viability and density can be determined before attempting to culture the protoplasts and induce cell wall regeneration. Applications include somatic hybridization through protoplast fusion to transfer genes between species.
Polymerase chain reaction powerpoint
The polymerase chain reaction (PCR) is a method used to rapidly amplify a specific region of DNA through repeated cycles of heating and cooling. It allows for the analysis and replication of short DNA or RNA sequences and is faster than using vectors, requiring only a small amount of target DNA. While useful for medical research, cloning, and forensic analysis, it is limited to short fragments under 5kb and requires prior knowledge of flanking sequences to design primers.
Protoplast culture
Somatic hybridization is a technique used to produce hybrid plants by fusing protoplasts (plant cells without cell walls) from two different plant species or varieties. There are several key steps:
1. Isolation of protoplasts from plant tissues using either mechanical or enzymatic methods. Enzymatic methods using cellulase and pectinase enzymes are more common.
2. Fusion of the protoplasts using chemical fusogens like polyethylene glycol (PEG) or physical methods like electrofusion. This results in hybrid cells called heterokaryons.
3. Selection and culture of the hybrid cells using techniques like antibiotic resistance or genetic markers.
4. Regeneration
POLYMERASE CHAIN REACTION
DNA amplification techniques include in vivo cloning and in vitro PCR. PCR was independently proposed in the 1970s and 1980s and allows selective amplification of DNA segments using a thermostable DNA polymerase. Key components of PCR include a template DNA, primers, DNA polymerase, nucleotides, and magnesium. During cycling, the DNA is denatured, primers anneal, and the polymerase extends the DNA. PCR has revolutionized molecular biology due to its ability to rapidly amplify specific DNA regions.
PCR
A detailed description about the basic steps involved in the - PCR - Polymerase Chain Reaction, its applications,its limitations and steps to overcome it.
Polymerase chain reaction
Polymerase chain reaction (PCR) is a technique used to amplify a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. PCR requires DNA polymerase, primers, nucleotides, buffer, and thermal cycling. It has many applications including detecting pathogens, DNA fingerprinting, and genetic testing.
PCR and its types
The document discusses the history and development of the polymerase chain reaction (PCR) technique. It describes how Kary Mullis invented PCR in 1985 and was awarded the Nobel Prize for it. It then explains the basic steps of PCR including denaturation, annealing of primers, and extension. Finally, it discusses several variations and applications of PCR including real-time PCR, asymmetric PCR, and comparisons to cloning techniques.
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Polymerase Chain Reaction
PCR introduction, history, enzymes in PCR, Reaction components,
PCR cycle, steps involved in PCR, RT-PCR, Applications.
PCR.ppt
PCR is a technique used to amplify a specific region of DNA across multiple cycles. It involves separating the DNA strands through heating, followed by primers annealing to the complementary DNA sequences. The DNA polymerase then extends the strands to exponentially increase copies of the target DNA. PCR has many applications in molecular biology, forensics, disease diagnosis and more due to its ability to amplify very small amounts of DNA.
Polymerase Chain Reaction(PCR)
Polymerase Chain Reaction (PCR) is a technique used to amplify small amounts of DNA sequences. It involves repeated cycles of heating and cooling of the DNA sample to denature and replicate the target DNA. Each cycle doubles the amount of target DNA, exponentially increasing its quantity for analysis. PCR uses primers, DNA polymerase, and dNTPs to selectively amplify the target DNA sequence. It has revolutionized molecular biology and is widely used for DNA cloning, detection of genetic diseases and mutations, forensic analysis, and more.
281 lec31 mol_tech3
PCR allows for the amplification of small amounts of DNA, generating millions of copies. It involves three steps - denaturation of the DNA template, annealing of primers, and extension of the primers by DNA polymerase. Repeating this process results in exponential growth in the number of DNA copies. DNA sequencing, such as the Sanger method, is used to determine the exact order of nucleotides in a DNA fragment and can be used to map genomes and detect genetic variations.
PCR.pdf
Polymerase chain reaction (abbreviated PCR) is a laboratory technique for rapidly producing (amplifying) millions to billions of copies of a specific segment of DNA, which can then be studied in greater detail.
PCR presentation
Consists of the main parts of PCR , type of PCR including the most advanced and most efficient.
It also includes history of PCR.
PCR is a machine used to make multiple copies of gene, while gene is a part of DNA. it has 3 steps , initiaition, elongation, termination. which required different temperature for different step. these slides includes most information about PCR.
Basics of Molecular Biology.ppt
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Polymerase Chain Reaction
Polymerase chain reaction (PCR) is a technique used to amplify a specific segment of DNA, generating millions of copies. It involves repeated cycles of heating and cooling DNA in the presence of DNA polymerase and primers to selectively amplify the target sequence. PCR is used in a wide range of applications including medical diagnosis, cloning, forensic analysis, and biological research.
Pcr
PCR is a technique used to amplify a specific DNA sequence. It involves repeated cycles of heating and cooling of the DNA sample to separate and copy the DNA strands. Key components of PCR include primers, DNA polymerase, and dNTPs. Variations of PCR allow for applications such as detecting gene expression, sequencing DNA, and quantifying DNA. Limitations include errors during amplification and potential contamination issues.
Pcr
Polymerase Chain Reaction (PCR) allows for targeted amplification of specific DNA sequences. PCR works by repeated cycles of heating and cooling of the DNA sample to denature the double strand into single strands and allow primers to anneal and new strands to extend. This process can generate billions of copies of the target sequence. Key components of PCR include DNA template, primers, Taq polymerase, nucleotides, and repeated thermal cycling. PCR has many applications in research and forensic analysis by enabling amplification of rare DNA sequences.
PCR
The document discusses polymerase chain reaction (PCR), a technique used to amplify specific DNA sequences. It describes the basic steps of PCR including denaturation of DNA, annealing of primers, and extension of DNA. Key requirements for PCR like DNA template, primers, DNA polymerase and buffers are explained. Commonly used DNA polymerases and their properties are outlined. Various applications and types of PCR including quantitative real-time PCR and reverse transcriptase PCR are summarized. The principles and methods of real-time PCR using DNA-binding dyes or fluorescent probes are briefly described.
Dna replication
DNA replication is the process by which DNA copies itself for transmission to daughter cells. In the late 1950s, three models were proposed for how DNA replicates: conservative, semi-conservative, and dispersive. Experiments showed that the semi-conservative model is correct, where each parental DNA strand acts as a template to replicate a new partner strand. DNA replication requires DNA and RNA polymerases, helicase, topoisomerases, primase, ligase and other proteins. It occurs through initiation, elongation and termination steps in both prokaryotes and eukaryotes, though eukaryotes have multiple replication origins and use RNA primers on the lagging strand.
Polymerase chain reaction
description of polymerase chain reaction and its principle and mechanism and requirements and applications and different types of pcr
Gene cloning and polymerase chain reaction
In these you are able to know about the gene cloning basic steps and Polymerase chain reaction process also there is an brief description about the ideal property shown by vectors which are lambda and M13 phases and there are lots of things in these slides
Polymerase Chain Reaction (PCR)
Polymerase chain reaction (PCR) is a technique developed by Kerry Mullis in 1984 that uses thermal cycling to amplify a specific DNA across several orders of magnitude, generating millions of copies of the target DNA segment. It involves repeated cycles of separating DNA strands through heating, annealing primers to the strands through cooling, and extending the primers with a thermostable DNA polymerase through heating. This allows for rapid and efficient amplification of targeted DNA regions.
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REPLICATION VS POLYMERASE CHAIN REACTION
- 2. REPLICATION • Replication occur in cell. • DNA replication is the copying of DNA. • It typically takes a cell just a few hours to copy all of its DNA. • DNA replication is semi-conservative (i.e. one strand of the DNA is used as the template for the growth of a new DNA strand). • This process occurs with very few errors (on average there is one error per 1 billion nucleotides copied). • More than a dozen enzymes and proteins participate in DNA replication.
- 4. ENZYME INVOLVED IN DNA REPLICATION • DNA Polymerase • DNA Ligase • Primase • Helicase • Topoisomerase • Single stranded binding protein
- 5. DEOXYRIBONUCLEIC ACID (DNA) • DNA is a double helix, made up of nucleotides. • NUCLEOTIDE – Pentose Sugar(Deoxyribose Sugar + Nitrogenous Base(A-T/G-C) + Phasphoric Acid • A nucleotide triphosphate is a 1 sugar + 1 base + 3 phosphates. • When a nucleoside triphosphate joins the DNA strand, two phosphates are removed.
- 6. DNA REPLICATION ENZYMES • Helicase untwists the two parallel DNA strands • Topoisomerase relieves the stress of this twisting • Single-strand binding protein binds to and stabilizes the unpaired DNA strands • Primase is the enzyme that can start an RNA chain from scratch and it creates a primer (a short stretch RNA with an available 3’ end) that DNA polymerase can add nucleotides to during replication. • DNA polymerase can only add nucleotides to 3’ end of growing strand. • One strand (referred to as the leading strand) of DNA is synthesized continuously and the other strand (referred to as the lagging strand) in synthesized in fragments (called Okazaki fragments) that are joined together by DNA ligase.
- 7. POLYMERASE CHAIN REACTION (PCR) • PCR means to amplify a particular piece of DNA . • PCR was invented in the 1984 by Kary mullis, as a way to make numerous copies of DNA fragments in the laboratory. • Polymerase chain reaction enables large amounts of DNA to be produced from very small samples (10nl) • There is a repeating cycle of separation of double DNA strands and synthesis of a complementary strand for each • It revolutionized biological methods specially in molecular cloning in a way that it has became an inseparable & irreplaceable part of molecular investigations.
- 9. COMPONENT OF PCR FOR PERFORM IN LABORATORY • DNA (your DNA of interest that contains the target sequence you wish to copy) • A heat-stable DNA Polymerase (like Taq Polymerase) • All four nucleotide triphosphates (ATP, GTP, TTP, CTP) • Buffers, MgCl2, Distilled water etc. • Two short, single-stranded DNA molecules that serve as primers • Thin walled tubes (PCR Tubes) • Thermo - cycler (a device that can change temperatures dramatically in a very short period of time)
- 10. THE MAIN STEPS OF PCR • The basis of PCR is temperature changes and the effect that these temperature changes have on the DNA. • In a PCR reaction, the following series of steps is repeated 20-40 times (Note: 25 cycles usually takes about 2 hours and amplifies the DNA fragment of interest 100,000 fold) Step 1: Denature DNA At 95C, the DNA is denatured (i.e. the two strands are separated) Step 2: Primers Anneal At 40C- 65C, the primers anneal (or bind to) their complementary sequences on the single strands of DNA Step 3: DNA polymerase Extends the DNA chain At 72C, DNA Polymerase extends the DNA chain by adding nucleotides to the 3’ ends of the primers.
- 11. PROCESS • Separation achieved by heating to 95oC – no suitable helicase • DNA polymerase can’t work on completely single stranded DNA – double stranded regions needed at the start of sequence to be copied: primers (short sequences DNA) complementary to bases at start of region to be copied used To synthesize primers , base sequence at start must be known TTAACGGGGCCCTTTAAA.....…TTTAAACCCGGGTTT AATTGCCCCGGGAAATTT.....…AAATTTGGGCCCAAA TTAACGGGGCCCTTTAAA.....…TTTAAACCCGGGTT AATTGCCCCGGGAAATTT.......................> and: <........................................TTTAAACCCGGGTTT AATTGCCCCGGGAAATTT........AAATTTGGGCCCAAA
- 12. • Pollution. • Poor precision. • Hard to get quantitative data • The most accurate & feasible technique to determine the amount & concentration of products. • Rapid cycling (30 minutes to 2 hours). • Specific & sensitive. • Not much more expensive. POLYMERASE CHAIN REACTIOON ADVANTAGE DISADVANTAGES