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If the DNA is cooled slowly, enough time is allowed for the bases on the ssDNA to re-allign and form the proper Watson/Crick base pairs. However, if the denaturation is followed by rapid cooling, there is not enough time for the nitrogenous bases to line up in an ordered fashion. In this rapid cooling case, the bases form random associations with nearby bases and the ssDNA does not re-assume an ordered double helix structure.
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Why do you keep competent cells in ice?
to avoid denaturation of the genetic material in this case DNA
Which DNA nucleotide pair is most stable?
GC pairing is most stable and require maximum energy to dissoicate. This is reason that rate of DNA denaturation depends upon the GC content of DNA.
Why heat DNA to 94 degrees?
depends what you intend to do, usually in a pcr reaction it denatures the DNA so you get 2 seperated strands
If E. coli DNA polymerase was used instead of thermus aquaticus DNA polymerase in a pcr polymerase chain reaction procedure then will reaction be carried out in waterbath?
no e coli DNA pol can not with stand high temperature if so for every denaturation step we must add fresh enzyme
What is pcr and its application?
PCR or polymerase chain reaction is a method to amplify a fragment of DNA. PCR reaction contains template DNA, primers, dNTPs, polymerase enzyme, buffer and water. The thermocycler manage the heat and time to synthesize DNA (denaturation, annealing and extension). The main application is one can amplify the gene or DNA of interest to millions of copies by using this prior to cloning.
Effect of hcl on DNA?
denaturation
How does high salt concentration influence denaturation kinetics of DNA?
High salt concentration slows the denaturation of DNA because it stabilizes the charges
Why initial denaturation step is done before denaturation step in PCR?
To make sure the double-strand DNA template is separated into single strands.
How do you destroy the hydrogen bonds holding DNA together?
It can be done by heating and the process is called denaturation of protein of DNA
Why extraction buffer is added for isolation of DNA?
Extraction Buffer is used to maintain pH of the solution.which prevents denaturation of DNA.
Why do you keep competent cells in ice?
to avoid denaturation of the genetic material in this case DNA
What is DNA cloning versus DNA re-naturing?
They are two different un related phenomena. In DNA cloninig, we cut a vector DNA and ligate our DNA of interest with the vector by DNA ligase, propagate the clones in E.coli or other host cells. DNA denaturation appears when you heat the DNA to higher temperature (above 60 degree Celsius). This can be reversed by cooling down the denatured DNA, where the two strands of DNA molecule will come closer and regain their NATIVE form by so called renaturation.
What is the mechanism of alkaline denaturation of dna?
Adding hydroxyl ions will deprive protons from DNA molecules that are needed to form a hydrogen bridge bond.
Why you give denaturation and neutralization to gel treatment?
For denaturation :-To eliminate hydrogen bonds with sodium hydroxide (NaOH)To denature double stranded DNA into single stranded DNAFor neutralization :-Neutralize the gel to get the pH that DNA can bind to the membrane.Destroy any remaining RNA present in sample
Which DNA nucleotide pair is most stable?
GC pairing is most stable and require maximum energy to dissoicate. This is reason that rate of DNA denaturation depends upon the GC content of DNA.
Why heat DNA to 94 degrees?
depends what you intend to do, usually in a pcr reaction it denatures the DNA so you get 2 seperated strands
If e. coli DNA polymerase was used instead of thermus aquaticus DNA polymerase in a pcr polymerase chain reaction procedure what would happen?
Unlike Taq DNA polymerase, E.coli DNA polymerase is not heat-stable and will denature during the strand denaturation step of the PCR reaction.
