I run a PCR with the sputum product which was microscopy positive, but after PCR the result was negative.
perhaps you have some troubles in tour extraction method
bwfore amplification you.can measure DNA through UV specrtrophotometer
Before DNA amplification you can measure the mass as well as the size by using DNA mass ladder and Size markers . These are simplest means to measure size and mass. however if you want to know precisely the amount of dna present per microlitre either you can go for NANO drop.
DNA mass ladders are specifically created for quantitative estimation of DNA mass in gels. These ladders consist of equimolar mixtures of DNA fragments for determining the mass of unknown DNA samples on gels in the low and high molecular weight ranges.
Double-stranded DNA size markers with band sizes ranging from 72 bp to 23.1 kb are available for use in agarose gel electrophoresis. These markers are suitable for estimation of fragment length.
After amplification you can ndo the same above and for further studies you can go for clloning and sequencing to authenticate your results
Have you done gel electrophoresis after extraction the DNA, and then quantified it. If there was no problem in these steps, the problem might be in your PCR protocol.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology