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peace upon you im working on 3 human gens about ( 200 ,300 , 400 )bp . my samples genamic DNA from blood . my supervisor dilut the primers to 100 pmol. i use Accua pwoer primix = 5 microliter total reaction volume = 20 microliter ( 9 microliter DNA ,, 1 microliter primers ) program : 95 c ..5min 95 c .. 1min 55 c ..1min 72 c ..1min 72 c .. 7 min 35 cycles (thermal cycler pcr ) at first the pcr is ok and the product perfect .... then primer dimer result and no product . what's going on ??? |
Re: peace upon you Hey there, You can try two things here: 1. Reduce the annealing temp. Rather setup a gradient. 2. Add only 1 primer, wait for about 5-6 cycles and then add the second primer. |
Re: peace upon you hi since it work at the first time : you have to check the step the amount of the genomic DNA you have try to increase the annealing tem 58 really you have check the technical error good luck :) |
Re: peace upon you thanx for your welcome |
Re: peace upon you thanks for response . |
Re: peace upon you that help me a lot. thanx for all of you |
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