Cot Curve_Dr. Sonia.pdf
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The document describes Cot curve analysis, a technique used to determine DNA sequence complexity in genomes. DNA is denatured by heating and allowed to renature at different time points. The percentage of single-stranded DNA is plotted against Cot value, the product of DNA concentration and renaturation time. Species with more complex genomes have higher Cot1/2 values as complementary sequences take longer to encounter each other. Eukaryotic genomes contain repetitive sequences that affect the Cot curve shape, with highly repetitive sequences renaturing fastest and single-copy sequences slowest.
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Eukaryotic DNA replication
Eukaryotic DNA replicationMarudhar Kesari Jain College for Women Vaniyambadi - 635 751, Tamil Nadu, INDIA.
The document summarizes eukaryotic DNA replication. It discusses that DNA replication in eukaryotes is more complex than prokaryotes due to larger genome size and chromatin packaging. The key stages of eukaryotic replication are similar to prokaryotes, including origin of replication, formation of replication forks, semiconservative replication and synthesis of leading and lagging strands. However, eukaryotic replication involves additional proteins and is slower due to chromatin remodeling required to access DNA.Transposable elements
Transportable elements are DNA Sequences that move from one location in a chromosome to another within the same chromosome or into another chromosome.
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These are also known as “Jumping genes”.
Sanger sequencing
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STRUCTURE AND ORGANIZATION OF CHROMATIN
This document provides information on the structure and organization of chromatin. It discusses three main levels of chromatin organization:
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2) The 30nm chromatin fiber, where nucleosomes are further compacted into a solenoid structure through histone H1 binding.
3) Higher-order compaction into metaphase chromosomes, where DNA is organized into loops anchored to a protein scaffold.
The document also describes different chromatin types (euchromatin and heterochromatin), chromatin composition, and models of chromatin structure including the nucleosome model.
Dna replication in eukaryotes
This document describes the process of DNA replication in eukaryotes. It occurs in S phase of the cell cycle and involves three main stages: initiation, formation of the initiation complex, and elongation. Initiation requires the assembly of pre-replication complexes containing ORC, Cdc6, Cdt1 and MCM proteins. In S phase, Cdc45 and GINS are recruited to form the initiation complex. Elongation proceeds bidirectionally from replication forks, with leading strand synthesis continuous and lagging strand discontinuous via Okazaki fragments. Replication terminates at telomeres.
Cot curve
Cot value and Cot Curve analysis is a technique for measuring DNA complexity based on renaturation kinetics. DNA is denatured and allowed to reanneal, with larger DNA taking longer. Cot value accounts for DNA concentration, time, and buffer effects, representing repetitive sequences - lower Cot means more repeats. Examples show bacteria have nearly all single-copy DNA, while mouse has varying proportions of single-copy, middle repetitive, and highly repetitive sequences. Cot curve analysis provides information on genome size, complexity, and proportions of sequence types.
Dna replication eukaryotes
DNA replication is the process by which DNA copies itself in living cells. It occurs in three main steps: initiation, elongation, and termination. Initiation begins at origins of replication, where proteins assemble into pre-replication complexes. During elongation, helicase unwinds the DNA strands and DNA polymerase adds complementary nucleotides to each strand. Termination occurs when the replication forks meet, with telomerase ensuring complete replication of chromosome ends.
FORMS OF DNA
The document discusses different forms of DNA structure that can be adopted based on environmental conditions. The main forms discussed are B-DNA, A-DNA, Z-DNA, C-DNA, D-DNA and E-DNA. B-DNA is the most common form, having a right-handed double helix structure with 10 base pairs per turn. A-DNA and Z-DNA are also double helical but have different structural characteristics than B-DNA such as base pair spacing and groove size. The various forms arise in response to changes in humidity, ionic conditions and DNA sequence composition.
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STRUCTURE AND ORGANIZATION OF CHROMATIN
This document provides information on the structure and organization of chromatin. It discusses three main levels of chromatin organization:
1) Nucleosomes, which involve DNA wrapping around histone proteins in "beads on a string" structures.
2) The 30nm chromatin fiber, where nucleosomes are further compacted into a solenoid structure through histone H1 binding.
3) Higher-order compaction into metaphase chromosomes, where DNA is organized into loops anchored to a protein scaffold.
The document also describes different chromatin types (euchromatin and heterochromatin), chromatin composition, and models of chromatin structure including the nucleosome model.
Dna replication in eukaryotes
This document describes the process of DNA replication in eukaryotes. It occurs in S phase of the cell cycle and involves three main stages: initiation, formation of the initiation complex, and elongation. Initiation requires the assembly of pre-replication complexes containing ORC, Cdc6, Cdt1 and MCM proteins. In S phase, Cdc45 and GINS are recruited to form the initiation complex. Elongation proceeds bidirectionally from replication forks, with leading strand synthesis continuous and lagging strand discontinuous via Okazaki fragments. Replication terminates at telomeres.
Cot curve
Cot value and Cot Curve analysis is a technique for measuring DNA complexity based on renaturation kinetics. DNA is denatured and allowed to reanneal, with larger DNA taking longer. Cot value accounts for DNA concentration, time, and buffer effects, representing repetitive sequences - lower Cot means more repeats. Examples show bacteria have nearly all single-copy DNA, while mouse has varying proportions of single-copy, middle repetitive, and highly repetitive sequences. Cot curve analysis provides information on genome size, complexity, and proportions of sequence types.
Dna replication eukaryotes
DNA replication is the process by which DNA copies itself in living cells. It occurs in three main steps: initiation, elongation, and termination. Initiation begins at origins of replication, where proteins assemble into pre-replication complexes. During elongation, helicase unwinds the DNA strands and DNA polymerase adds complementary nucleotides to each strand. Termination occurs when the replication forks meet, with telomerase ensuring complete replication of chromosome ends.
FORMS OF DNA
The document discusses different forms of DNA structure that can be adopted based on environmental conditions. The main forms discussed are B-DNA, A-DNA, Z-DNA, C-DNA, D-DNA and E-DNA. B-DNA is the most common form, having a right-handed double helix structure with 10 base pairs per turn. A-DNA and Z-DNA are also double helical but have different structural characteristics than B-DNA such as base pair spacing and groove size. The various forms arise in response to changes in humidity, ionic conditions and DNA sequence composition.
Nucleosomes
Nucleosomes are the fundamental repeating subunits of eukaryotic chromatin that package DNA into a compact structure. They are composed of 146 base pairs of DNA wrapped around an octamer of histone proteins, resembling beads on a string. This represents the first order of DNA compaction. Higher orders of compaction involve the nucleosomes winding further to form solenoid fibers, scaffold loops, chromatids, and finally full chromosomes. Nucleosomes allow the long DNA molecules to fit within cell nuclei while also regulating genetic expression.
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Semiconservative replication
Meselson and Stahl conducted an experiment using E. coli bacteria to test the hypothesis that DNA replicates semi-conservatively. They grew the bacteria in medium containing a heavy isotope of nitrogen, then switched the bacteria to medium with a light isotope. Analysis of DNA densities over multiple generations provided evidence that DNA replication results in one old strand and one new strand in each daughter molecule, supporting the semi-conservative model.
Retrotransposons
Retrotransposons are genetic elements that copy and paste themselves throughout the genome using an RNA intermediate and reverse transcription. There are two main types: LTR retrotransposons, which mimic retroviruses through reverse transcription of an RNA copy into DNA; and non-LTR retrotransposons like LINEs and SINEs. LINEs (Long Interspersed Nuclear Elements) are autonomous retrotransposons over 6kb with endonuclease and reverse transcriptase proteins. SINEs (Short Interspersed Nuclear Elements) are shorter than 300bp and non-autonomous, relying on LINEs to reverse transcribe themselves.
Transcription in prokaryotes and eukaryotes
Transcription is the process of synthesizing RNA from DNA and involves four main stages - initiation, elongation, termination, and post-transcription processing. It occurs differently in prokaryotes and eukaryotes. In prokaryotes, transcription and translation are coupled and occur in the cytoplasm, while in eukaryotes transcription occurs separately in the nucleus. The document provides details on the mechanisms and factors involved in each stage of transcription for both prokaryotes and eukaryotes.
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This document discusses cryptic satellite DNA. It begins by introducing satellite DNA as highly repetitive sequences that form distinct bands during density gradient centrifugation. Cryptic satellite DNA does not form distinct bands due to base methylation. The document then provides two examples of cryptic satellite DNA: 1) In Drosophila virilis, there are three major satellites and a cryptic one. 2) In a hermit crab, one major very highly repeated DNA and three minor variants comprising inverted repeats account for 30% of its genome. These minor variants are considered cryptic satellites.
Dna topology
DNA topology refers to the coiling and knotting of DNA strands. DNA exists in vivo in a negatively supercoiled state, with the strands intertwined about each other. Enzymes called topoisomerases regulate and maintain the appropriate level of supercoiling by introducing transient breaks into DNA strands to allow strand passage and control overwinding or underwinding of the double helix.
DNA Denaturation and Renaturation, Cot curves
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- Renaturation is the spontaneous rewinding of single strands back into the original double helix structure when denaturing conditions are removed, through base pairing of complementary strands.
- C0t curves plot the fraction of single strands renatured versus the product of DNA concentration and time, and can indicate the complexity and size of the original DNA sample based on renaturation rates. More complex DNA with more dissimilar sequences takes longer to renature
Second genetic code overlapping and split genes
1) Eukaryotic genes can be organized in complex ways, including overlapping genes where coding sequences partially overlap, and split genes where coding sequences are interrupted by non-coding intron sequences.
2) Overlapping genes were discovered in bacteriophage X174, where the coding sequences of genes D and E overlap but are translated in different reading frames.
3) Split genes have exons, which are the coding sequences included in mRNA, and introns, which are intervening non-coding sequences not included in mRNA. Split genes were first observed in animal viruses in 1977.
Nucleic acid sequencing- introduction,DNA sequencing
DNA sequencing is a fundamental process in biotechnology that determines the order of nucleotides in a DNA molecule. There are two main methods: the chemical method developed by Maxam and Gilbert, and the enzymatic method developed by Sanger and Coulson. DNA sequencing involves degrading DNA fragments into smaller pieces, sequencing the individual fragments, and ordering the fragments to determine the full DNA sequence.
TRANSCRIPTION IN EUKARYOTES
Eukaryotic transcription is carried out in the nucleus of the cell and proceeds in three sequential stages: initiation, elongation, and termination. Eukaryotes require transcription factors to first bind to the promoter region and then help recruit the appropriate polymerase.
Eukaryotic Chromosome Organisation
This document discusses eukaryotic chromosome organization. It notes that eukaryotic cells contain many chromosomes in the nucleus, with each species having a characteristic number. Chromosomes are made up of DNA and proteins like histones. DNA is wrapped around histones to form structures called nucleosomes, which are further compacted through multiple levels of coiling and folding involving other proteins. This allows the long DNA molecules to fit within cell nuclei.
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DNA replication in prokaryotes involves initiation, elongation, and termination phases. Initiation begins with the binding of initiator proteins to the origin of replication, unwinding the DNA helix to form replication forks. Elongation synthesizes the leading and lagging strands bidirectionally away from the origin using DNA polymerases. Termination occurs when the replication forks meet, completing duplication of the chromosome.
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This document discusses the central dogma of biology and the process of transcription. It describes the three main steps of transcription - initiation, elongation, and termination. Initiation involves the RNA polymerase binding to the promoter sequence on DNA and separating the DNA strands to form an open complex. Elongation is the addition of nucleotides to synthesize RNA. Termination can occur via either Rho-independent or Rho-dependent mechanisms, with the former utilizing a terminator sequence and hairpin structure in the RNA and the latter involving the Rho protein.
Eukaryotic promoters
Eukaryotic promoters are DNA sequences located upstream of genes that recruit transcriptional machinery to initiate gene transcription. Promoters can range from 100-1000 base pairs in length and are located near transcription start sites on the same DNA strand in the 5' region. Promoters serve as binding sites for RNA polymerase and help initiate transcription of downstream DNA. The strength of a promoter can be regulated by the presence or absence of binding proteins. There are different types of promoters including constitutive, tissue-specific, synthetic, and inducible promoters. Methods like DNase footprint assays and online prediction tools can determine promoter sites.
Holliday junction Model by kk
INTRODUCTION
TERMENOLOGY
SITES OF HOLLIDAY JUNCTION
HOMOLOGOUS RECOMBINATION
HOLLIDAY MODEL
DOUBEL STRAND BREAK MODEL
IN PROKAYOTIC RECOMBINATION
IN EUKARYOTIC RECOMBINATION
SITE SPECIFIC RECOMBINATION
SERINE RECOMBINATION
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TRANSPOSITION
ILLEGITIMATE RECOMBINATION
CONCLUSION
REFRENCES
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STEPS INVOLVE IN TRANSCRIPTION.
RNA POLYMERASES.
HISTORY OF RNA POLYMERASES.
STRUCTURE OF RNA POLYMERASES.
SUB UNITS OF RNA POLYMERASES.
TYPES OF RNA POLYMERASES.
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TRANSCRIPTION FACTORS INVOLVE IN EUKARYOTIC TRANSCRIPTION.
CONCLUSION.
REFERENCES.
Presenatation On Restriction Enzyme
Restriction enzymes are proteins that cut DNA at specific recognition sequences. There are three main types of restriction enzymes. Type I enzymes require cofactors and have three subunits. Type II enzymes recognize palindromic sequences and are the most commonly used. They cut DNA into blunt or sticky ends. Type III enzymes recognize two separate sequences and cut further from the recognition site. Restriction enzymes are important tools in biotechnology and DNA manipulation.
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Site specific recombination
Site-specific recombination involves DNA strand exchange between segments with sequence homology, mediated by site-specific recombinases (SSRs). SSRs recognize and bind to short DNA sequences, cleaving the DNA backbone to exchange helices and rejoin strands. They are classified into tyrosine and serine recombinase families based on mechanism. Tyrosine recombinases like Cre and Flp cleave DNA strands staggered by 6-8bp, linking DNA ends to the recombinase. Serine recombinases simultaneously cleave all four strands staggered by 2bp via phosphoserine bonds. SSRs have applications like tracking cell lineage, ablating genes, and inducing gene expression at specific developmental times.
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DNA polymerases are a group of enzymes that are used to make copies of DNA templates, essentially used in DNA replication mechanisms. These enzymes make new copies of DNA from existing templates and also function by repairing the synthesized DNA to prevent mutations. DNA polymerase catalyzes the formation of the phosphodiester bond which makes up the backbone of DNA molecules. It uses a magnesium ion in catalytic activity to balance the charge from the phosphate group.
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Meselson and Stahl conducted an experiment using E. coli bacteria to test the hypothesis that DNA replicates semi-conservatively. They grew the bacteria in medium containing a heavy isotope of nitrogen, then switched the bacteria to medium with a light isotope. Analysis of DNA densities over multiple generations provided evidence that DNA replication results in one old strand and one new strand in each daughter molecule, supporting the semi-conservative model.
Retrotransposons
Retrotransposons are genetic elements that copy and paste themselves throughout the genome using an RNA intermediate and reverse transcription. There are two main types: LTR retrotransposons, which mimic retroviruses through reverse transcription of an RNA copy into DNA; and non-LTR retrotransposons like LINEs and SINEs. LINEs (Long Interspersed Nuclear Elements) are autonomous retrotransposons over 6kb with endonuclease and reverse transcriptase proteins. SINEs (Short Interspersed Nuclear Elements) are shorter than 300bp and non-autonomous, relying on LINEs to reverse transcribe themselves.
Transcription in prokaryotes and eukaryotes
Transcription is the process of synthesizing RNA from DNA and involves four main stages - initiation, elongation, termination, and post-transcription processing. It occurs differently in prokaryotes and eukaryotes. In prokaryotes, transcription and translation are coupled and occur in the cytoplasm, while in eukaryotes transcription occurs separately in the nucleus. The document provides details on the mechanisms and factors involved in each stage of transcription for both prokaryotes and eukaryotes.
cryptic satellite DNA
This document discusses cryptic satellite DNA. It begins by introducing satellite DNA as highly repetitive sequences that form distinct bands during density gradient centrifugation. Cryptic satellite DNA does not form distinct bands due to base methylation. The document then provides two examples of cryptic satellite DNA: 1) In Drosophila virilis, there are three major satellites and a cryptic one. 2) In a hermit crab, one major very highly repeated DNA and three minor variants comprising inverted repeats account for 30% of its genome. These minor variants are considered cryptic satellites.
Dna topology
DNA topology refers to the coiling and knotting of DNA strands. DNA exists in vivo in a negatively supercoiled state, with the strands intertwined about each other. Enzymes called topoisomerases regulate and maintain the appropriate level of supercoiling by introducing transient breaks into DNA strands to allow strand passage and control overwinding or underwinding of the double helix.
DNA Denaturation and Renaturation, Cot curves
The document discusses DNA denaturation and renaturation, including:
- Denaturation involves unwinding the DNA double helix into single strands through heating or chemical treatment, disrupting hydrogen bonds between base pairs. This increases UV absorption.
- Renaturation is the spontaneous rewinding of single strands back into the original double helix structure when denaturing conditions are removed, through base pairing of complementary strands.
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Second genetic code overlapping and split genes
1) Eukaryotic genes can be organized in complex ways, including overlapping genes where coding sequences partially overlap, and split genes where coding sequences are interrupted by non-coding intron sequences.
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TRANSCRIPTION IN EUKARYOTES
Eukaryotic transcription is carried out in the nucleus of the cell and proceeds in three sequential stages: initiation, elongation, and termination. Eukaryotes require transcription factors to first bind to the promoter region and then help recruit the appropriate polymerase.
Eukaryotic Chromosome Organisation
This document discusses eukaryotic chromosome organization. It notes that eukaryotic cells contain many chromosomes in the nucleus, with each species having a characteristic number. Chromosomes are made up of DNA and proteins like histones. DNA is wrapped around histones to form structures called nucleosomes, which are further compacted through multiple levels of coiling and folding involving other proteins. This allows the long DNA molecules to fit within cell nuclei.
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DNA replication in prokaryotes involves initiation, elongation, and termination phases. Initiation begins with the binding of initiator proteins to the origin of replication, unwinding the DNA helix to form replication forks. Elongation synthesizes the leading and lagging strands bidirectionally away from the origin using DNA polymerases. Termination occurs when the replication forks meet, completing duplication of the chromosome.
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This document discusses the central dogma of biology and the process of transcription. It describes the three main steps of transcription - initiation, elongation, and termination. Initiation involves the RNA polymerase binding to the promoter sequence on DNA and separating the DNA strands to form an open complex. Elongation is the addition of nucleotides to synthesize RNA. Termination can occur via either Rho-independent or Rho-dependent mechanisms, with the former utilizing a terminator sequence and hairpin structure in the RNA and the latter involving the Rho protein.
Eukaryotic promoters
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Holliday junction Model by kk
INTRODUCTION
TERMENOLOGY
SITES OF HOLLIDAY JUNCTION
HOMOLOGOUS RECOMBINATION
HOLLIDAY MODEL
DOUBEL STRAND BREAK MODEL
IN PROKAYOTIC RECOMBINATION
IN EUKARYOTIC RECOMBINATION
SITE SPECIFIC RECOMBINATION
SERINE RECOMBINATION
TYROSINE RECOMBINATION
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CONCLUSION
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STEPS INVOLVE IN TRANSCRIPTION.
RNA POLYMERASES.
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SUB UNITS OF RNA POLYMERASES.
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TRANSCRIPTION FACTORS INVOLVE IN EUKARYOTIC TRANSCRIPTION.
CONCLUSION.
REFERENCES.
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DNA is the hereditary material found in humans and other organisms. It is a double-stranded molecule made up of nucleotides with four bases - adenine, guanine, cytosine, and thymine. Genes are sections of DNA that code for proteins or RNA chains and hold the information to build and maintain organisms. Vectors are DNA molecules used to transfer genes between cells. Common vectors include plasmids and viruses. The process of cloning involves isolating DNA from cells, cutting it into pieces using restriction enzymes, reassembling the pieces into a vector, and inserting the vector back into host cells.
C value, Cot Curve & Rot Curve L1-3.pdf
The lecture describes the basic concepts of C-value, Cot curve and Rot curve analysis, MCQ questions regarding the same. Queries are always welcome.... Dr. Nitin Wahi (wahink@gmail.com).
0.PDF
Prokaryotic genetic material differs from eukaryotes in several key ways:
1. Prokaryotes lack a membrane-bound nucleus and have their DNA located in the nucleoid. They typically have a single circular chromosome while eukaryotes have multiple linear chromosomes.
2. Prokaryotic genes are arranged in operons and expressed together, whereas eukaryotic genes each have their own promoter and are independently expressed.
3. DNA replication in prokaryotes is rapid and ongoing, starting from a single origin of replication site, while eukaryotes tightly regulate replication during the cell cycle.
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DNA replication requires many enzymes to accurately duplicate the genome. Key enzymes include DNA helicase which unwinds the DNA double helix, DNA polymerase which synthesizes new DNA strands using existing strands as templates, DNA ligase which seals DNA fragments after replication, and primase which synthesizes RNA primers for DNA polymerase to begin DNA synthesis. Accurate replication of DNA is essential for organisms to reproduce and pass genetic information between generations.
DNA Damage and DNA Repair- Dr. Sonia Angeline
DNA is constantly damaged by environmental and metabolic factors. The three main types of DNA damage are oxidative damage from reactive oxygen species, alkylation of bases by environmental chemicals, and hydrolytic deamination of bases. Cells have multiple DNA repair pathways to fix different types of lesions. These include base excision repair for single base changes, nucleotide excision repair for distortions to the DNA structure, mismatch repair for replication errors, and double-strand break repair using non-homologous end joining or homologous recombination. The SOS response is also activated in bacteria to induce DNA repair genes in response to damage.
Enzymes involved in homologous recombination.pdf
1. Key enzymes involved in homologous recombination in E. coli include RecBCD, RecA, RuvA, RuvB, and RuvC. RecBCD and RecA help initiate recombination by processing DNA and catalyzing strand exchange. RuvA, RuvB, and RuvC resolve Holliday junction intermediates through branch migration and resolution.
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Recombination Models.pdf
This document discusses molecular genetics topics including sex determination, genetic recombination, and transposons. It outlines four units of study: 1) Sex determination and dosage compensation, 2) Genetic recombination mechanisms and models, 3) Enzymes involved in homologous and site-specific recombination, and 4) Bacterial and eukaryotic transposons. The second section provides detailed descriptions of homologous recombination models including Holliday, Whitehouse, Meselson-Radding, and double-strand break repair pathways. Key enzymes in E. coli recombination such as RecBCD, RecA, RuvAB, and RuvC are also summarized.
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The document discusses human fetal sex determination and its implications in India. It provides background on India's preference for male children and the history of female infanticide. It summarizes the Prenatal Diagnostic Techniques Act passed in 1994 that banned determining the gender of fetuses to address the issue. The act was later upgraded in 2003 as the PC-PNDT Act to further ban related advertisements and technologies. However, sex-selective abortions still illegally occur and the act has been criticized by some. The document also describes medical techniques like amniocentesis and CVS that can determine fetal sex but also carry risks.
CHROMOSOME BANDING PATTERN_Dr. Sonia.pdf
This document discusses various chromosome banding techniques used to identify chromosomes and detect abnormalities. It describes:
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2) Banding techniques that selectively stain regions of chromosomes based on DNA composition, allowing identification. These include Q, G, R, C, and T bands.
3) The techniques involve staining with fluorescent dyes or Giemsa followed by heat/chemical treatments to differentially darken bands for analysis under microscopes. Banding patterns are consistent and chromosome-specific.
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The document discusses hybridoma technology for producing monoclonal antibodies. It describes the process which involves fusing antibody-producing B cells from immunized mice with myeloma tumor cells, using a chemical or electrical method. The resulting hybridoma cells are selected and cultured, retaining the antibody-producing ability of B cells and indefinite lifespan of tumor cells. The monoclonal antibodies produced can be extracted and purified for therapeutic and research applications.
Transgenic Animals.pdf
Transgenic animals are animals whose genome has been altered by the addition of foreign DNA from other species. This can be done to improve genetic traits like increased milk or wool production. Common methods to create transgenic animals include microinjecting DNA into fertilized eggs, using viruses to insert DNA, or manipulating embryonic stem cells. Transgenic mice and sheep are useful for research, as disease models, and to produce pharmaceuticals or other proteins.
Genome organization
The document discusses various aspects of genome organization, including:
1. Chromatin assembly begins with the incorporation of histone proteins to form nucleosomes, which are then folded and organized into higher order structures within the nucleus.
2. Genes can be split, overlapping, or pseudogenes. Split genes contain introns that are spliced out, while overlapping genes share nucleotide sequences. Pseudogenes are non-functional copies of genes.
3. Gene families consist of genes related by common ancestry that may be clustered or dispersed throughout the genome. Members can vary in sequence but often retain similar functions.
Gene transfer mechanisms
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- Later experiments by Avery, MacLeod and McCarty identified DNA as the molecule responsible for bacterial transformation. They showed that DNA is the genetic material that is able to transform one type of bacteria into another through uptake and incorporation of foreign DNA.
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X ray diffraction
X-ray diffraction is a technique that uses X-rays to determine the atomic and molecular structure of crystals. When X-rays hit a crystal, they cause the atoms to diffract into specific patterns determined by Bragg's law. By analyzing these diffraction patterns, information about the crystal structure such as lattice parameters and spacing between atomic planes can be determined. Common applications of XRD include identifying materials and determining their purity, structure, and properties.
Transmission electron microscope
The document discusses the transmission electron microscope (TEM). It begins by explaining that a TEM uses a beam of electrons rather than light to produce highly magnified images of thin specimens. It then provides details on the history and development of the TEM. The body of the document describes the main components of a TEM, including the electron gun, image producing system, and image recording system. It explains how each component works and its role in producing a magnified image. Applications of the TEM in fields like biology and nanotechnology are also mentioned.
Growth factors
The document discusses several growth factors including epidermal growth factor (EGF), fibroblast growth factor (FGF), platelet-derived growth factor (PDGF), and erythropoietin. It describes how these growth factors function to regulate cell growth, differentiation, and survival by binding to cell surface receptors and activating intracellular signaling pathways. Specific examples of their roles in wound healing, angiogenesis, and fetal development are provided.
ABT Introduction
This document provides information about animal biotechnology through several sections. It begins with an introduction that discusses the long history of animal biotechnology including traditional breeding techniques dating back to 5000 BC. It then covers the history of animal biotechnology from the 1970s to the present day, highlighting important milestones. Several sections follow on the scope, applications, and terminology of animal biotechnology including transgenic animals, cloning, animal models in research, vaccines, nutrition, and embryo transfer. The document concludes by defining common terminology used in animal cell culture.
GC and HPLC
Gas chromatography and high performance liquid chromatography are analytical techniques used to separate compounds in a mixture. GC uses an inert gas as the mobile phase to carry vaporized analytes through a column coated with a stationary phase for separation. HPLC forces a liquid mobile phase at high pressure through a column packed with porous particles to separate compounds based on interactions with the stationary phase. Both techniques separate components by differences in partitioning between the mobile and stationary phases, with detectors then identifying and quantifying the separated analytes.
Electrophoresis
1. Electrophoresis is a technique used to separate molecules like DNA, RNA, and proteins based on their size and electrical charge. An electric current is applied to move the molecules through a gel.
2. Smaller molecules move faster through the gel's pores than larger molecules. Electrophoresis conditions can be adjusted to separate molecules in a desired size range.
3. During electrophoresis, charged molecules migrate toward the electrode of opposite charge. Positively charged molecules move toward the cathode, while negatively charged molecules move toward the anode.
Bio occupational hazards
1) Occupational biohazards refer to infectious agents or hazardous biological materials that can harm workers' health directly through infection or indirectly by damaging the working environment.
2) Biological hazards in the workplace include viruses, bacteria, fungi, toxins, and other microorganisms that can enter the body through inhalation, contact, or ingestion and cause disease.
3) Controlling biological hazards involves proper use of personal protective equipment, good hygiene practices, sterilization procedures, and maintaining biosafety levels in laboratories commensurate with the pathogenicity of the microorganisms being handled.
Air sampling devices
The document discusses several air sampling devices used to collect airborne particles such as microorganisms, pollen, and spores. The slit sampler uses a rotating petri dish under a slit to directly impinge particles onto agar media. The Anderson sampler uses progressively smaller perforations to increase air speed and momentum to collect particles of different sizes on agar plates. The Burkard spore trap is a volumetric sampler that uses an adhesive tape to continuously trap particles over 24 hours to 1 week to analyze pollen and spore levels over time.
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Cot Curve_Dr. Sonia.pdf
- 1. Cot Curve Dr. M. Sonia Angeline KJC
- 2. Analysis of DNA Sequences in Eukaryotic Genomes The technique that is used to determine the sequence complexity of any genome involves the denaturation and renaturation of DNA. DNA is denatured by heating which melts the H-bonds and renders the DNA single- stranded. If the DNA is rapidly cooled, the DNA remains single-stranded. But if the DNA is allowed to cool slowly, sequences that are complementary will find each other and eventually base pair again. The rate at which the DNA reanneals (another term for renature) is a function of the species from which the DNA was isolated.
- 3. Analysis of DNA Sequences in Eukaryotic Genomes The Y-axis is the percent of the DNA that remains single stranded. This is expressed as a ratio of the concentration of single-stranded DNA (C) to the total concentration of the starting DNA (Co). The X-axis is a log-scale of the product of the initial concentration of DNA (in moles/liter) multiplied by length of time the reaction proceeded (in seconds). The designation for this value is Cot and is called the "Cot" value. The curve itself is called a "Cot" curve. As can be seen the curve is rather smooth which indicates that reannealing occurs slowing but gradually over a period of time. One particular value that is useful is Cot½ , the Cot value where half of the DNA has reannealed.
- 4. Cot= DNA Concentration (moles/L) X Renaturation time in sec X Buffer factor (Constant) Co- Concentration of DNA t- time taken for renaturation
- 5. Steps Involved in DNA Denaturation and Renaturation Experiments 1. Shear the DNA to a size of about 400 bp. 2. Denature the DNA by heating to 100oC. 3. Slowly cool and take samples at different time intervals. 4. Determine the % single-stranded DNA at each time point. The shape of a "Cot" curve for a given species is a function of two factors: 1. the size or complexity of the genome; and 2. the amount of repetitive DNA within the genome
- 6. If we plot the "Cot" curves of the genome of three species such as bacteriophage lambda, E. coli and yeast we will see that they have the same shape, but the Cot½ of the yeast will be largest, E. coli next and lambda smallest. Physically, the larger the genome size the longer it will take for any one sequence to encounter its complementary sequence in the solution. This is because two complementary sequences must encounter each other before they can pair. The more complex the genome, that is the more unique sequences that are available, the longer it will take for any two complementary sequences to encounter each other and pair. Given similar concentrations in solution, it will then take a more complex species longer to reach Cot½ .
- 8. Repeated DNA sequences, DNA sequences that are found more than once in the genome of the species, have distinctive effects on "Cot" curves. If a specific sequence is represented twice in the genome it will have two complementary sequences to pair with and as such will have a Cot value half as large as a sequence represented only once in the genome. Eukaryotic genomes actually have a wide array of sequences that are represented at different levels of repetition. Single copy sequences are found once or a few times in the genome. Many of the sequences which encode functional genes fall into this class. Middle repetitive DNA are found from 10s - 1000 times in the genome. Examples of these would include rRNA and tRNA genes and storage proteins in plants such as corn. Middle repetitive DNA can vary from 100-300 bp to 5000 bp and can be dispersed throughout the genome. The most abundant sequences are found in the highly repetitive DNA class. These sequences are found from 100,000 to 1 million times in the genome and can range in size from a few to several hundred bases in length. These sequences are found in regions of the chromosome such as heterochromatin, centromeres and telomeres and tend to be arranged as a tandem repeats.
- 9. These sequences are found in regions of the chromosome such as heterochromatin, centromeres and telomeres and tend to be arranged as a tandem repeats. The following is an example of a tandemly repeated sequence: ATTATA ATTATA ATTATA // ATTATA Genomes that contain these different classes of sequences reanneal in a different manner than genomes with only single copy sequences. Instead of having a single smooth "Cot" curve, three distinct curves can be seen, each representing a different repetition class. The first sequences to reanneal are the highly repetitive sequences because so many copies of them exist in the genome, and because they have a low sequence complexity. The second portion of the genome to reanneal is the middle repetitive DNA, and the final portion to reanneal is the single copy DNA.
- 10. The following diagram depicts the "Cot" curve for a "typical" eukaryotic genome