a PCR and gel electrophoresis was carried out to obtain the results. they are negative jak2 v617f samples (ref genome - NG_009904 )
Your primers are annealing elsewhere on the genome. Run an annealing temperature gradient and at higher annealing temperatures the non specific bands should disappear. Alternatively run a sample with 1%, 2% etc up to 8% DMSO at any single annealing temperature that works without dmso which will make the primer annealing more stringent until at too high concentration the pcr will fail but before that concentration the product will be much cleaner. Both of these techniques rely on increased stringency of the primer either at higher temperature or higher dmso
Can you explain more briefly about your problem?
From what I understood, this gel is after PCR but they are negative i.e. expected bands are not there. Looking at the gel it looks like you have plasmids in the gel or if it is genomic DNA then your primers are not specific and have multiple binding sites which are resulting in these non-specific amplifications. Therefore, you need to check your primers or change the programme of your PCR.
Divya Das I have taken samples of 5 jak2 v617f negative patients (lanes 11 - 15) and lane 16 is a control (water). I have run a pcr and got these results.. I'm trying to find rare mutations by performing a sangar sequencing afterward.
My issue is that I'm trying to figure out the reason for these non specific bands
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