I have used dreamtaq and amplitaq with both plain and nested PCR but it does not seem to work really well, The image is really smeared with a lot of non specific binding when extracted on 1.5% Agarose ?
a picture of a gel ,the primer sequences and the pcr conditions would be very helpful in answering this question please Saqib as it may be the pcr and not the dna quality which is the problem
Have your specific primers worked to amplify the ssDNA of parvovirus before? Are they the correct sense? Do your ladders or markers appear properly in the gel? What are your positive and negative controls? And how do they appear in the gel?
Hi,
These primers are specific for canine parvovirus. Please modify thermal cycles 94/2min + 40 (98/ 10sec+50/30sec +72/1 min) and 72/ 5min and keep 4 degree celcilus. I used 0.75 ul primer (10mM).
Good luck
Dung
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