I am doing Newcastle Disease Virus PCR, the bands are visible but always on the bottom of the agarose gel and lower than 100 bp even with the positive control and the dna ladder show up perfectly. The RNA nanodrop is around 150-200 picomol. I used 1% agarose gel with 4 microliter amplicon. Electrophoresis is 100 v for 60 minutes. I also tried several annealing temperature but the bands are still at the bottom of the gel.
Giorgia Pertile
Anastasia Irawan how much is in theory your PCR-product? But your template is DNA or cDNA (RNA)?
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Giorgia Pertile
Anastasia Irawan Do you performed the primer with DNA before test it on cDNA?
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