I used to run gel before with no problems, but lately I started to get samples missing. I do prepare new TBE buffer everytime I ran a gel, but this seems not to be the problem, since I get other bands appears to be fine in the same gel.
How did you checked RNA integrity from PCR products?? All samples from same template and same primerS?
If template id different maybe that particular gene is not amplified from that particular template.
I checked RNA purity after RNA extraction, then I did PCR, yes samples from same template and primers. I used the same primer for all the samples
Dear Mustafa,
If you do not see any band for some samples, the first question is : did the PCR work for those samples ? I assume not, or you would see at least one band per sample.
Dear Vincent,
What I am doing is just repeating an experiment used to work well for me with exactly same conditions (primers pairs, cDNA, gel conc, PCR program). I also tried cleaning pipettes and bench with RnaseZAP from sigma. What I am going to do is further purification for my RNA samples and do RT and PCR and see how it goes.
Thanks
Did u used RNA for PCR? or synthesised cDNA ffrom RNA first? as you mentioned that you used RNA for PCR.
In my opinion there is only one reason if you have missing bands in a gel : there is no PCR product in the corresponding lane(s), simple as that !
I also agree with Vincent's ans or maybe you forgot to add something in that particular pcr tube reaction.
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