I have a problem with conventional PCR electrophoresis, we are analyzing the deletion of ccr5 delta 32. And this material started to cause problems recently, there was no change in the protocol and before it worked normally. All reagents have already been changed, we are using conventional master mix.
What can it be?
what exactly is the problem You are running the gel too high a voltage causing some smearing and you have primer dimer that would probably benefit from using a hot start enzyme so the only problem that I can see is that you look to have over amplification of your samples. Where is the no template control on this picture because it may be that you have pcr contamination in your pcr and any amplification of the negative control would indicate this.
Looks like lots of bands on that gel. Are they not the expected size (hard to tell since they are so smeared/smiling)? Or are you getting contamination (band in negative)?
You can solve the smearing by also loading less sample & making sure the gel rig equipment is clean & has new running buffer. Salts can also cause smears.
can you give more information as to the type of problem you are having? No product, different product?
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