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Old 05-16-2014, 04:11 PM   #2
Jafar Jabbari
Location: Melbourne

Join Date: Jan 2013
Posts: 1,238

This indicates that there is a primer heterodimer with strong bonds. First step would be to raise annealing temperature to favour your intended amplification while reducing unwanted primer-dimer interaction. If that is not helpful, next step would be to design new region specific primer(s) and analyse primer-primer interaction between your amplification primers (primers included with appended sequences) carefully to ensure that there is little interaction in your intended annealing temperature. You will need to take into account the salt concentration in your enzyme buffer as well.
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