How does a higher annealing temperature affect the specificity of a PCR reaction? - annealing temperature calculator
I really do not understand this problem in my mission forensics. If someone enlighten us on this, it would be greatly appreciated.
Sunday, November 8, 2009
Annealing Temperature Calculator How Does A Higher Annealing Temperature Affect The Specificity Of A PCR Reaction?
3 comments:
Hello,
Therefore, PCR in a series of steps to be divided. The initial denaturation, annealing of primers, then the extension.
During the annealing step to bind the primer to the complementary sequence. The higher the temperature the more specific the union. The annealing temperature (Ta) depends on the length and composition of the primer and is typically ~ 5'c below the MT.
In PCR, the annealing temperature a "landing" is very high at the beginning and then gradually decreases with each cycle. This means that you soon will have the right things and when the temperature drops the remains, ie areas with a single database of my games, so the ability to bind.
Of course you do not want the annealing temperature is too high, primers will have different timing needs perfectly demonstrated in the analysis of a series of primers temperature gradient to achieve the best results (narrow band did not know if it's on a gel shown).
Hope this helps
Hello,
Therefore, PCR in a series of steps to be divided. The initial denaturation, annealing of primers, then the extension.
During the annealing step to bind the primer to the complementary sequence. The higher the temperature the more specific the union. The annealing temperature (Ta) depends on the length and composition of the primer and is typically ~ 5'c below the MT.
In PCR, the annealing temperature a "landing" is very high at the beginning and then gradually decreases with each cycle. This means that you soon will have the right things and when the temperature drops the remains, ie areas with a single database of my games, so the ability to bind.
Of course you do not want the annealing temperature is too high, primers will have different timing needs perfectly demonstrated in the analysis of a series of primers temperature gradient to achieve the best results (narrow band did not know if it's on a gel shown).
Hope this helps
Hello,
Therefore, PCR in a series of steps to be divided. The initial denaturation, annealing of primers, then the extension.
During the annealing step to bind the primer to the complementary sequence. The higher the temperature the more specific the union. The annealing temperature (Ta) depends on the length and composition of the primer and is typically ~ 5'c below the MT.
In PCR, the annealing temperature a "landing" is very high at the beginning and then gradually decreases with each cycle. This means that you soon will have the right things and when the temperature drops the remains, ie areas with a single database of my games, so the ability to bind.
Of course you do not want the annealing temperature is too high, primers will have different timing needs perfectly demonstrated in the analysis of a series of primers temperature gradient to achieve the best results (narrow band did not know if it's on a gel shown).
Hope this helps
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