When I used gp5 and gp6 primers my product length was 150 bp ,according that i got it bands, but even i got band in non template control. After then i use everything fresh (pcr mix ,water) even though i got it bands in NTC ,how to resolve my problem?
Deepthi your problem seems to be with cross contamination, not with the primers. gp5/6 is a good primer pair. If you have changed all your reagents and still getting amplicons in the NTC then you must think about your pipettes and the clean air cabinet that you are using. are you using same pipettes for DNA extraction and PCR set-up? or making the PCR master mix outside a clean air cabinet, or in the same lab where you extract DNA or analyze DNA? These questions need to be satisfactorily answered before switching to a different primer pair.
best wishes,
SD
Thank you sir .
yes i using same pipettes for DNA extraction
so these time i will use new pipettes and determine accuracy of NTC
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