After PCR,many PCR product(bants) were observed in the agrose electrophoresis. Although we have modified the Tm, primer and template concentration .
Probably the change of primer concentration let them to amplify aspecific sequences with the same nucleotide pattern but different length/weight.
I think your primers were more concentrated. you can try again by diluting the forward and reverse primers.
What is you downstream application? If you need o clone the product you can cut it out from gel (assuming there is a sufficient size difference between the desired product and the others).
Hi, I think we need more information that we can help you properly. Cheers, Nadine
Dear Jun,
Their are 'n' number of probabilities for multiple banding pattern in PCR product.
If you are looking for the single band i.e. specific amplicon, then something is wrong in your protocol. I will suggest to recheck your chemicals, primers, always keep negative control to rule out contamination possibility.
All the best.
Hi,
I had this problem before. I guess, in addition primer concentration, annealing tempreture, etc, its better to check your Agarose gel (Percentage, Contamination,...) and Power instrument.
good luck
This could be possible chance of contamination, improper primer design (Forming dimers) or technical error. The best to rectify use negative control, check once again your primer design and use 10um for PCR. If still you see unspecific band try HOT START TECHNIQUE, prepare your PCR reaction on ICE and place on pre-heated PCR machince.
Thank You.
Without more information there is less chance to help you properly. Add some more information about your PCR etc.
I want to take the target gene about 1-2Kb from the vector. There are many nonspecfic bands in the electrophresis. I have some adjustment on primer concentration, template contration. It is very surprised that I found many times in my lab, enve if small fragment will be amplied.
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