Improve Research Reproducibility A Bio-protocol resource

What is the effect of high annealing temperatures?

John Khalil Oct 20, 2022

Hello, I recently cloned a gene with a gradient annealing temperature, the results show that when the annealing temperature is 67 ° C and above, the gene bands in gel electrophoresis will be clearer, but I am worried that the high annealing temperature will bring some bad effects, does Read more Read less more less

follow or recommend this question

685 Views

Answer

Recommend

1 answer

Sort by: most helpful / most recent

Sharon Akanbi Answered Oct 21, 2022

University of Leeds

The double strands of DNA are mainly linked by hydrogen bonds. The higher the annealing temperature, the greater the thermal energy to break the hydrogen bond, and the less likely it is for the primers to combine with the template, so that the amplified bands will be less, and sometimes there will be no bands. However, only primers that match the base of the template to a high degree can be combined at higher annealing temperatures, and the specificity of the resulting bands will be higher.

15 helpful

0 unhelpful

0 comments down

My answer

Write your answer...

References (optional)

add Add more

Post an Answer