Usually I am doing PCR with 0.5 ul of Taq polymerase (abm) per reaction. Since I wanted to increase the sensitivity of target amplification, I tried PCR with 1.0 ul of Taq. It gave some bands in confirmed healthy samples which were with correct size as in the positive control. What would be the reason for that?
(The target I am amplifying is Leishmania in DNA extracted from patients' bone marrow materials.)
I think you need to make sure the concentration of extracted DNA
regards
I think you need to make sure the concentration of extracted DNA
regards
How was the result of PCR using 0.5 ul of Taq polymerase?
It might be both too low concentration of Taq that might cause false negative or non-specific band that sequencing should be applied for confirming.
Sounds like you have contamination in your reactions. Do you have a good positive control sample? Try again with a full set of controls (positive control DNA, DNA without Leishmania, and a water control).
Hi Bhagya
You tried for touch down PCR?
touch down PCR will increase the specificity.
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