Dna denaturation and renaturation
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DNA denaturation refers to the melting of double-stranded DNA into single strands by breaking the hydrogen bonds between bases. It can be caused by elevated temperature, extremes of pH, or non-physiological salt concentrations. Denatured DNA is characterized by increased absorbance and decreased viscosity. DNA renaturation is the reformation of the double helix structure when denatured DNA is placed under conditions that allow the strands to reanneal through base pairing. These processes of denaturation and renaturation are important techniques used to understand genome size, genetic relatedness, and repetitive sequences.
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DNA replication in eukaryotes
DNA replication is the process by which a cell makes an identical copy of its DNA. It occurs in three main steps: initiation, elongation, and termination. Initiation begins at origins of replication and involves unwinding of the DNA double helix by helicases. During elongation, DNA polymerases add nucleotides to build new strands based on the existing DNA templates. Termination occurs when the replication forks meet at the end of the DNA molecule. DNA replication is semi-conservative, meaning each new DNA molecule contains one original and one new strand of DNA.
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.
Prokaryotic DNA replication
DNA replication in prokaryotes begins with the unwinding of DNA at the origin of replication by enzymes like DnaA and DnaB helicase. This produces two replication forks that move in opposite directions. The leading strand is replicated continuously while the lagging strand is replicated discontinuously in short segments called Okazaki fragments. DNA polymerase III is the main enzyme that synthesizes new DNA. Replication terminates at the terminus region when the DnaB helicase is stopped by protein Tus bound to Ter sequences.
Denaturation and renaturation of dna
DNA is a double-helix molecule that carries genetic instructions. It is composed of two strands of polynucleotides made up of nucleotides, each containing a nitrogenous base, sugar, and phosphate. The strands are stabilized by hydrogen bonds between complementary bases and base-stacking interactions. DNA can be denatured into single strands by elevated temperature, extreme pH, low salt concentrations, or chemicals that disrupt hydrogen bonding between strands. Denaturation temperature depends on factors like base composition and length. Renaturation occurs when double-stranded DNA is cooled under conditions that allow the strands to re-form hydrogen bonds and complementary base pairing.
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.
A, b, and z forms of dna
There are three main forms of DNA structure: A-DNA, B-DNA, and Z-DNA. B-DNA is the most common form found under physiological conditions, having a right-handed double helix with 10.5 base pairs per turn. A-DNA forms under dehydrating conditions and has a wider helix with 11 base pairs per turn. Z-DNA is a left-handed helix that forms with alternating purine-pyrimidine sequences, containing 12 base pairs per turn in a narrow, zig-zag structure. While B-DNA is most prevalent, the structure can vary depending on sequence and environmental conditions.
Structure of DNA
The present slide describes the DNA Structures and the components of the DNA along with the illustrative structures.
DNA Denaturation
it is a little powerpoint presentation by me about DNA DEnaturation wish you all benefit from it .
for any , contact me at iahmedaly55@gmail.com
Recommended
DNA replication in eukaryotes
DNA replication is the process by which a cell makes an identical copy of its DNA. It occurs in three main steps: initiation, elongation, and termination. Initiation begins at origins of replication and involves unwinding of the DNA double helix by helicases. During elongation, DNA polymerases add nucleotides to build new strands based on the existing DNA templates. Termination occurs when the replication forks meet at the end of the DNA molecule. DNA replication is semi-conservative, meaning each new DNA molecule contains one original and one new strand of DNA.
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.
Prokaryotic DNA replication
DNA replication in prokaryotes begins with the unwinding of DNA at the origin of replication by enzymes like DnaA and DnaB helicase. This produces two replication forks that move in opposite directions. The leading strand is replicated continuously while the lagging strand is replicated discontinuously in short segments called Okazaki fragments. DNA polymerase III is the main enzyme that synthesizes new DNA. Replication terminates at the terminus region when the DnaB helicase is stopped by protein Tus bound to Ter sequences.
Denaturation and renaturation of dna
DNA is a double-helix molecule that carries genetic instructions. It is composed of two strands of polynucleotides made up of nucleotides, each containing a nitrogenous base, sugar, and phosphate. The strands are stabilized by hydrogen bonds between complementary bases and base-stacking interactions. DNA can be denatured into single strands by elevated temperature, extreme pH, low salt concentrations, or chemicals that disrupt hydrogen bonding between strands. Denaturation temperature depends on factors like base composition and length. Renaturation occurs when double-stranded DNA is cooled under conditions that allow the strands to re-form hydrogen bonds and complementary base pairing.
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.
A, b, and z forms of dna
There are three main forms of DNA structure: A-DNA, B-DNA, and Z-DNA. B-DNA is the most common form found under physiological conditions, having a right-handed double helix with 10.5 base pairs per turn. A-DNA forms under dehydrating conditions and has a wider helix with 11 base pairs per turn. Z-DNA is a left-handed helix that forms with alternating purine-pyrimidine sequences, containing 12 base pairs per turn in a narrow, zig-zag structure. While B-DNA is most prevalent, the structure can vary depending on sequence and environmental conditions.
Structure of DNA
The present slide describes the DNA Structures and the components of the DNA along with the illustrative structures.
DNA Denaturation
it is a little powerpoint presentation by me about DNA DEnaturation wish you all benefit from it .
for any , contact me at iahmedaly55@gmail.com
Dna replication in prokaryotes
DNA replication in prokaryotes occurs through bidirectional replication from an origin of replication. It involves three main steps - initiation, elongation, and termination. During initiation, helicase unwinds the DNA at the origin of replication, forming a replication fork. In elongation, DNA polymerase extends RNA primers to synthesize new leading and lagging strands. The lagging strand is synthesized discontinuously in short Okazaki fragments. Termination occurs when replication is complete and the replicated circular DNA molecules are separated by topoisomerase II.
Various model of DNA replication
1. There are four main models of DNA replication: rolling circle replication, theta replication, bidirectional replication of linear DNA, and telomere replication.
2. Rolling circle replication involves nicking circular DNA and using one strand as a template to produce multiple copies of the original circular DNA.
3. Theta replication occurs in prokaryotes and involves unwinding circular DNA at an origin of replication and replicating bi-directionally to form a theta-shaped structure.
4. Bidirectional replication of linear DNA involves unwinding DNA at origins of replication and using leading and lagging strand synthesis to replicate in both directions until the ends of the linear genome are reached.
DNA damage and_repair
DNA damage and causes along with their various Repair mechanisms. Slideshare suitable for students of Class 12, B.Sc. and M.Sc.
Physiochemical properties of dna
This document discusses several physiochemical properties of DNA including:
- Absorption of UV light at 260nm, which can be used to determine DNA concentration. Double stranded DNA absorbs less light than single stranded DNA.
- Buoyant density of 1.73 g/cm3 in cesium chloride, which can estimate GC content through centrifugation separation. Higher GC DNA migrates lower.
- Denaturation of the double helix into single strands above 90°C or through chemical means like low pH or urea, monitored by increased UV absorption.
- Renaturation of single strands back into the double helix upon cooling through hydrogen bonding of complementary bases.
Telomerase replication
Telomers & Telomerase replication.
THIS WAS MY PRESENTATION TOPIC IN CLASS. THOUGHT OF SHARING IT AND HOPE IT HELPS.
Sos repair
SOS response was discovered by Miroslav Radman. It's a part of DNA repair system- synthesizes enzymes required for DNA repair. Cellular response to UV damage.
Sos repair mechanism
SOS repair
a system that repairs severely damaged bases in DNA by base excision and replacement, even if there is no template to guide base selection. This process is a last resort for repair and is often the cause of mutations.
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.
- 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
Dna binding proteins
The document discusses DNA binding proteins. It describes how DNA is wrapped around histone proteins to form nucleosomes, which resemble "beads on a string". There are five main types of histone proteins - H1, H2A, H2B, H3, and H4. Histone proteins can be modified through processes like acetylation and methylation, which affect gene expression. Other non-histone proteins use motifs like zinc fingers and helix-turn-helix to bind DNA in a sequence-specific manner and regulate transcription.
Origin of replication, replication fork, enzymes
The document summarizes DNA replication in three main points:
1) DNA replication involves the semi-conservative duplication of DNA in which each new DNA molecule contains one old and one new strand. It is facilitated by various enzymes and occurs through multiple steps.
2) Replication initiates at the origin of replication and proceeds bidirectionally, forming a replication fork. The leading strand is replicated continuously while the lagging strand forms discontinuous Okazaki fragments.
3) Key enzymes involved include DNA polymerases, helicase, primase, ligase, topoisomerases and single-strand binding proteins which help unwind, copy and join DNA strands for accurate duplication of genetic material.
Dna replication in eukaryotes
DNA replication in eukaryotes proceeds in a coordinated process of initiation, elongation, and termination. Initiation begins at specific origin of replication sequences and involves the assembly of pre-replication complexes. During elongation, helicases unwind the DNA strands exposing templates for DNA polymerase enzymes to synthesize new daughter strands. Termination occurs when the polymerases reach the end regions called telomeres, which are then extended by the enzyme telomerase to allow replication to continue.
Transcription in eukaryotes
transcription process in eukaryotes their factors RNA polymerase enzyme an intiation elongation termination process
DNA Replication -
1. DNA replication begins with the unwinding of the DNA double helix by the helicase enzyme. This exposes the two strands of DNA to serve as templates for DNA synthesis.
2. RNA primers are synthesized by the primase enzyme and DNA polymerase adds complementary nucleotides to the 3' end of each primer, elongating it into DNA.
3. The leading strand is synthesized continuously in the 5' to 3' direction while the lagging strand is synthesized in fragments called Okazaki fragments which are later joined by DNA ligase.
4. Several enzymes are involved including DNA polymerase, ligase, helicase, topoisomerase and others to accurately and efficiently copy DNA. DNA replication ensures faithful transmission of
Different Forms of DNA
The following slides contains a brief comparison of the different forms of the DNA. It includes A-DNA, B-DNA , and Z-DNA.
It also briefs about the conditions that would favor the transition from one form to the another
Recombination
recombination
types of recombination
models of recombination
biological roles of recombination
repetitive and non repetitive dna.pptx
1. Eukaryotic DNA contains repetitive and non-repetitive segments. Repetitive DNA makes up around 50% of the human genome and consists of sequences that are present in copies numbering over a million.
2. Repetitive DNA is divided into highly, moderately, and uniquely repetitive sequences based on copy number. Highly repetitive sequences are present in over 100,000 copies and include satellite and centromeric DNA. Moderately repetitive sequences have between 100-10,000 copies, like ribosomal RNA genes.
3. Non-repetitive or unique sequences make up around 50% of the human genome and contain protein-coding genes and other sequences required for gene expression that generally exist in only
Dna replication in prokaryotes
Introduction to Replication, Replication process in Prokaryotes
Proposed models of replication,
Semi-conservative method by meselson and sthal
Enzymes of DNA replication
Enzymes of DNA replicationMarudhar Kesari Jain College for Women Vaniyambadi - 635 751, Tamil Nadu, INDIA.
DNA replication is the process where one original DNA molecule is copied to produce two identical DNA molecules. It occurs through semiconservative replication where each strand of the original DNA serves as a template for the production of the complementary strand. The replication fork forms during DNA replication through the unwinding of the DNA double helix by helicases. It has two branching prongs that serve as templates for the leading and lagging strands which are synthesized by DNA polymerase.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.
DNA Replication in eukaryotes and prokaryotes
1. DNA replication is the process by which daughter DNA molecules are synthesized from a parental DNA template. It ensures the genetic information is transferred to the next generation with high fidelity.
2. Replication occurs semi-conservatively such that each new double helix contains one strand from the original parent DNA and one newly synthesized strand. It also occurs bidirectionally from an origin of replication.
3. DNA polymerases are the key enzymes that catalyze DNA synthesis. Other important enzymes and proteins include primase, helicase, topoisomerase, ligase, and single-stranded DNA binding proteins. Together they facilitate the initiation, elongation and termination of DNA replication.
denaturation and renaturation of dna.pptx
denaturation and renaturation of dna.pptxDepartment of Biotechnology, Kamaraj college of engineering and technology
DNA is a double-helix molecule that carries genetic instructions. It is composed of two strands called polynucleotides made up of nucleotides, each containing a nucleobase (A, T, C, or G), sugar, and phosphate. The strands are stabilized by hydrogen bonds between nucleotides and base stacking. DNA can be denatured by heat, pH extremes, or chemicals, breaking the hydrogen bonds and separating the strands. Denaturation temperature depends on factors like composition, length, and environment. Renaturation occurs when strands reconnect under appropriate conditions.Denaturation of DNA
DNA is a double-helix molecule that carries genetic instructions. It is composed of two strands of polynucleotides made up of nucleotides, each containing a nitrogenous base, sugar, and phosphate. The strands are stabilized by hydrogen bonds between complementary bases and base-stacking interactions. DNA can be denatured into single strands by elevated temperature, extreme pH, low salt concentrations, or chemicals that disrupt hydrogen bonding between strands. Denaturation temperature depends on factors like base composition and length. Renaturation occurs when double-stranded DNA is cooled under conditions that allow the strands to re-form hydrogen bonds and complementary base pairing.
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Dna replication in prokaryotes
DNA replication in prokaryotes occurs through bidirectional replication from an origin of replication. It involves three main steps - initiation, elongation, and termination. During initiation, helicase unwinds the DNA at the origin of replication, forming a replication fork. In elongation, DNA polymerase extends RNA primers to synthesize new leading and lagging strands. The lagging strand is synthesized discontinuously in short Okazaki fragments. Termination occurs when replication is complete and the replicated circular DNA molecules are separated by topoisomerase II.
Various model of DNA replication
1. There are four main models of DNA replication: rolling circle replication, theta replication, bidirectional replication of linear DNA, and telomere replication.
2. Rolling circle replication involves nicking circular DNA and using one strand as a template to produce multiple copies of the original circular DNA.
3. Theta replication occurs in prokaryotes and involves unwinding circular DNA at an origin of replication and replicating bi-directionally to form a theta-shaped structure.
4. Bidirectional replication of linear DNA involves unwinding DNA at origins of replication and using leading and lagging strand synthesis to replicate in both directions until the ends of the linear genome are reached.
DNA damage and_repair
DNA damage and causes along with their various Repair mechanisms. Slideshare suitable for students of Class 12, B.Sc. and M.Sc.
Physiochemical properties of dna
This document discusses several physiochemical properties of DNA including:
- Absorption of UV light at 260nm, which can be used to determine DNA concentration. Double stranded DNA absorbs less light than single stranded DNA.
- Buoyant density of 1.73 g/cm3 in cesium chloride, which can estimate GC content through centrifugation separation. Higher GC DNA migrates lower.
- Denaturation of the double helix into single strands above 90°C or through chemical means like low pH or urea, monitored by increased UV absorption.
- Renaturation of single strands back into the double helix upon cooling through hydrogen bonding of complementary bases.
Telomerase replication
Telomers & Telomerase replication.
THIS WAS MY PRESENTATION TOPIC IN CLASS. THOUGHT OF SHARING IT AND HOPE IT HELPS.
Sos repair
SOS response was discovered by Miroslav Radman. It's a part of DNA repair system- synthesizes enzymes required for DNA repair. Cellular response to UV damage.
Sos repair mechanism
SOS repair
a system that repairs severely damaged bases in DNA by base excision and replacement, even if there is no template to guide base selection. This process is a last resort for repair and is often the cause of mutations.
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.
- 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
Dna binding proteins
The document discusses DNA binding proteins. It describes how DNA is wrapped around histone proteins to form nucleosomes, which resemble "beads on a string". There are five main types of histone proteins - H1, H2A, H2B, H3, and H4. Histone proteins can be modified through processes like acetylation and methylation, which affect gene expression. Other non-histone proteins use motifs like zinc fingers and helix-turn-helix to bind DNA in a sequence-specific manner and regulate transcription.
Origin of replication, replication fork, enzymes
The document summarizes DNA replication in three main points:
1) DNA replication involves the semi-conservative duplication of DNA in which each new DNA molecule contains one old and one new strand. It is facilitated by various enzymes and occurs through multiple steps.
2) Replication initiates at the origin of replication and proceeds bidirectionally, forming a replication fork. The leading strand is replicated continuously while the lagging strand forms discontinuous Okazaki fragments.
3) Key enzymes involved include DNA polymerases, helicase, primase, ligase, topoisomerases and single-strand binding proteins which help unwind, copy and join DNA strands for accurate duplication of genetic material.
Dna replication in eukaryotes
DNA replication in eukaryotes proceeds in a coordinated process of initiation, elongation, and termination. Initiation begins at specific origin of replication sequences and involves the assembly of pre-replication complexes. During elongation, helicases unwind the DNA strands exposing templates for DNA polymerase enzymes to synthesize new daughter strands. Termination occurs when the polymerases reach the end regions called telomeres, which are then extended by the enzyme telomerase to allow replication to continue.
Transcription in eukaryotes
transcription process in eukaryotes their factors RNA polymerase enzyme an intiation elongation termination process
DNA Replication -
1. DNA replication begins with the unwinding of the DNA double helix by the helicase enzyme. This exposes the two strands of DNA to serve as templates for DNA synthesis.
2. RNA primers are synthesized by the primase enzyme and DNA polymerase adds complementary nucleotides to the 3' end of each primer, elongating it into DNA.
3. The leading strand is synthesized continuously in the 5' to 3' direction while the lagging strand is synthesized in fragments called Okazaki fragments which are later joined by DNA ligase.
4. Several enzymes are involved including DNA polymerase, ligase, helicase, topoisomerase and others to accurately and efficiently copy DNA. DNA replication ensures faithful transmission of
Different Forms of DNA
The following slides contains a brief comparison of the different forms of the DNA. It includes A-DNA, B-DNA , and Z-DNA.
It also briefs about the conditions that would favor the transition from one form to the another
Recombination
recombination
types of recombination
models of recombination
biological roles of recombination
repetitive and non repetitive dna.pptx
1. Eukaryotic DNA contains repetitive and non-repetitive segments. Repetitive DNA makes up around 50% of the human genome and consists of sequences that are present in copies numbering over a million.
2. Repetitive DNA is divided into highly, moderately, and uniquely repetitive sequences based on copy number. Highly repetitive sequences are present in over 100,000 copies and include satellite and centromeric DNA. Moderately repetitive sequences have between 100-10,000 copies, like ribosomal RNA genes.
3. Non-repetitive or unique sequences make up around 50% of the human genome and contain protein-coding genes and other sequences required for gene expression that generally exist in only
Dna replication in prokaryotes
Introduction to Replication, Replication process in Prokaryotes
Proposed models of replication,
Semi-conservative method by meselson and sthal
Enzymes of DNA replication
Enzymes of DNA replicationMarudhar Kesari Jain College for Women Vaniyambadi - 635 751, Tamil Nadu, INDIA.
DNA replication is the process where one original DNA molecule is copied to produce two identical DNA molecules. It occurs through semiconservative replication where each strand of the original DNA serves as a template for the production of the complementary strand. The replication fork forms during DNA replication through the unwinding of the DNA double helix by helicases. It has two branching prongs that serve as templates for the leading and lagging strands which are synthesized by DNA polymerase.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.
DNA Replication in eukaryotes and prokaryotes
1. DNA replication is the process by which daughter DNA molecules are synthesized from a parental DNA template. It ensures the genetic information is transferred to the next generation with high fidelity.
2. Replication occurs semi-conservatively such that each new double helix contains one strand from the original parent DNA and one newly synthesized strand. It also occurs bidirectionally from an origin of replication.
3. DNA polymerases are the key enzymes that catalyze DNA synthesis. Other important enzymes and proteins include primase, helicase, topoisomerase, ligase, and single-stranded DNA binding proteins. Together they facilitate the initiation, elongation and termination of DNA replication.
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Dna denaturation and renaturation
- 1. ZOOLOGY, JIWAJI UNIVERSITY, GWALIOR 1 TOPIC:- DNA: DENATURATION AND RENATURATION Dr. Ramkumar Lodhi Conservation Biology Lab ramkumarlodhi73@gmail.com
- 2. SYNOPSIS Introduction:- What is DNA DNA denaturation Causes Elevated temperature Extremes of pH Non physiological salts Characters of denatured DNA DNA renaturation Applications Conclusion References 2
- 3. INTRODUCTION DNA is a molecule composed of 2 strands running in opposite directions to form a double helix structure carrying genetic information. Each strands composed of de-oxy ribose sugar molecules linked together by phosphate linkage. The two strands are held together along their length with hydrogen bonds between the bases. 3
- 4. DNA DENATURATION 4 DNA denaturation refers to the melting of doubled stranded DNA to generate two single strands by breaking the hydrogen bonds.
- 5. CAUSES OF DENATURATION Denaturation can occur when nucleic acids are subjected to: elevated temperature extremes of pH non-physiological concentrations of salt, organic solvents, urea, or other chemical agents. 5
- 6. Elevated temperature The most common type of denaturation is thermal denaturation Tm: Temperature at which DNA is half denatured. • Above Tm DNA is single stranded • Below Tm DNA is double stranded 6
- 7. Extremes of pH At high pH, the hydroxide ions (negatively charged ions) can pull hydrogen ions from base pairs (forming H bond between two strands) causing them to separate. High pH will alter the charged groups that are involved in H- bond H-bond cannot occur at pH>11.3 and DNA denatures 7
- 8. Non physiological salts Low salt concentration could also denature DNA double strands by removing ions that stabilize the negative charges on the two strands from each other. The Tm value is lowered by the addition of Urea. In 8M urea, Tm is decreased by nearly 20 degree celsius DNA can be denatured with 95% formamide at room temperature only. 8
- 9. Characters of Denatured DNA Hyperchromic: increase the absorbance(A260) upon denaturation The rate of increase in absorbance is directly proportional to the rate of denaturation Viscosity decreases upon denaturation 9
- 10. DNA Renaturation Renaturation or annealing: is the formation of base repairs and complementary strands of DNA come back together. Renaturation occurs if double stranded DNA is heated above Tm then the temperature is slowly decreased under appropriate conditions 10
- 12. Applications : Understanding the genome size and complexity. Understanding genetic relatedness. Understanding relative proportion of repetitive sequences. 12
- 13. Conclusions These processes form the basis of many important techniques for DNA manipulating. Though there are many techniques associated with DNA denaturation, the end result is the same that is to break the bonds between the strands and new molecules are formed which can then be compared as desired. The ideal process of DNA denaturation depends on what the DNA needs to be use for, how accurate and specific the comparisons need to be and the volume of the material that has to be processed. 13
- 14. REFERENCES:- Orby’s illustrated reviews of biochemistry Lippincott’s illustrated reviews of biochemistry www.biologydiscussion.com Harper's biochemistry 5th edition Review lecture on "illustrated biochemistry “ 14
- 15. 15