Recently we have been working on serotyping Salmonella by using traditional PCR. We do the runs on three phases. Phase 1 to confirm the microbr. Phase 2 to identify the somatic cell and phase 3 is divided to identify the two different phases of flagella. We use several primers for this work. Unfortunatelly, we were not able to get any result except for one run for phase 1. This successful run was achived by diluting the primer, where we normally use some volume directly from the primer stock. Later on we tried to do phase 1 run with the same dilution again but again it did not work. We d not centrifuge upon the preparation of pcr mix. We use distilled water for the the lypholised primers.
What could be the reason for this issue??
PCR not working
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