I have a small oligo primer of 20 mer and the PCR product size should be at 158bp and i am only getting bands at 1.5kb band on gel electrophoresis which is too large!! My oligo primers (F and R) have the same melting temperature at 62'C so i used an annealing temperature equal to the melting temperature for 28 cycles. Can anyone help me on how to calculate the required temperatures and how to programme my elongation?????????????
Can you please send me a link to help me with PCR ??????????
thanx in advance.
Edited by youyou, 20 May 2009 - 01:50 AM.