Hello, I have a question about primer length.
What if the primer becomes shorter about 8~10 mer and set annealing temperature between 25~30 ℃ ?
Will there be problem in proceeding PCR?
Thank you.
Why and how do you know your primer length and annealing temperature are shortened?
Dear Yoonho
It is generally accepted that the optimal length of PCR primers is 18-22 bp. This length is long enough for adequate specificity and short enough for primers to bind easily to the template at the annealing temperature. Primers that are too short would anneal non-specifically at several positions on a long DNA template, resulting in non-specific copies. Also, too low temperature annealing may possibly lead to non-specific products caused by a high number of base pair mismatches.
Hello Yoonho,
The recommended length for primer would be 18-20mers and if you go below that you may end up with non-specific bindings.
However, if it is unavoidable, try to increase the GC content if possible.
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