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Denaturation

Definition

noun

(1) A process in which the folding structure of a protein is altered due to exposure to certain chemical or physical factors (e.g. heat, acid, solvents, etc.), causing the protein to become biologically inactive.

(2) A process in which the structure of nucleic acid is disrupted, such as the dissociation of a double stranded DNA into a single stranded state by heating.

(3) The process of making food or drink (e.g. alcohol) unfit for human consumption by the deliberate addition of a noxious substance.


Supplement

Biological proteins (such as enzymes) unfold and lose their active state when exposed to denaturing agents (e.g. strong acids or bases, heat, solvents, and salts). This is crucial especially when enzymes lose their structure and function as catalysts. The substrates can no longer bind to the active site, and biochemical process is therefore disrupted. The proteins can regain their natural active state if the denaturing agent is removed. However, there are instances in which the process is irreversible.


Related forms: denature (verb), denatured (adjective).

Related term: alkali denaturation test


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Annealing Temparature

... obviously contain added restriction sites and the matching part is quite short. I would anneal the first cycle at 50°C and the rest at 65°C. Denaturation for 10s will be enough, but a bit higher temperature 95-98°C might be better. Elongation depends on polymerase, so 1 min can be enough ...

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by vladCH
Sat Feb 22, 2014 7:54 pm
 
Forum: Molecular Biology
Topic: Annealing Temparature
Replies: 4
Views: 1362

Annealing Temparature

First, you seem to have big difference in the Tms, second the Tms seem to be quite high (62 - 69°C). Are you sure your denaturation in the beginning is only 1 min or is it a typo? Why do you have such long denaturation during cycling? Why do you have such long annealing? What polymerase ...

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by JackBean
Tue Feb 18, 2014 12:40 pm
 
Forum: Molecular Biology
Topic: Annealing Temparature
Replies: 4
Views: 1362

Calculating the molecular concentration of DNA

Concentration of 2 ng/ml of dsDNA will still be 2 ng/ml of ssDNA after denaturation. Concentration of 2 mmol/ml of dsDNA will be 4 mmol/ml of ssDNA after denaturation.

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by JackBean
Mon Oct 08, 2012 2:07 pm
 
Forum: Molecular Biology
Topic: Calculating the molecular concentration of DNA
Replies: 6
Views: 4962

Re: Help - Why can't DNA be parallel?

Nice question :) Unfortunately, 3D picture would not help here. I was thinking, why couldn't DNA anneal in parallel configuration after denaturation in vitro . The answer is pretty simple - because it is not capable to do so. Look on this picture http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/B/BasePairing.gif ...

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by keetner
Tue Oct 02, 2012 9:07 pm
 
Forum: Molecular Biology
Topic: Help - Why can't DNA be parallel?
Replies: 7
Views: 4965

Re: Help - Why can't DNA be parallel?

Nice question :) Unfortunately, 3D picture would not help here. I was thinking, why couldn't DNA anneal in parallel configuration after denaturation in vitro . The answer is pretty simple - because it is not capable to do so. Look on this picture http://users.rcn.com/jkimball.ma.ultranet/BiologyPages/B/BasePairing.gif ...

See entire post
by JackBean
Sun Sep 30, 2012 10:01 pm
 
Forum: Molecular Biology
Topic: Help - Why can't DNA be parallel?
Replies: 7
Views: 4965
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