The PCR product of Wheat I run over the gel was intact in the wells of and not run over the gel while some of these wells were but of them were not.....can anybody tell me the reason behind it? what is the problem?
If you could post a picture of the gel, it might help to identify the problem. If I understand you correctly, there is material staying just in/around the wells of your gel, but no PCR product observed lower in the gel. Usually if you have material that does not move through the gel, it is either not DNA or it is genomic DNA template and your PCR reaction has failed.
Thanks alot Sir Mr. Eric, I think this pic will give you an idea to sort out my problem.....
If you are using genomic DNA as your template, I would say the material in the wells is genomic DNA which is usually far too large to migrate through a gel made for PCR. This will not stop PCR product from running through the gel though. You do not see your PCR product, because it did not amplify. There can be many reasons for failing to amplify. Have you successfully amplified this product before?
Mr.Eric, yes I have used this Genomic DNA for many other genes but few of them have given such type of result. As far as my knowledge is concern either the genomic DNA is highly concentrated or PCR product is not well amplified. Anyhow, thanks alot for your feedback and cooperation.
Hi Muhammad, I met a similar situation as you were. I amplified my plenti-plasmid, which is 12K. The PCR product should be 10K. I only put 1 ng template. But I get many products in the wells, while the 10K product cannot see. Did you solve your problem? Do you have any suggestions? Thank you so much in advance for any advice.
Attached is the picture for your reference. Reaction 1,2,3 is the same plasmid but with different primers. The 2 and 3 are the right size, but I don't know why reaction 1 is not run in the gel.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- DNA