I was using High Pure PCR Template Preparation Kit version 20 from Roche to isolate DNA from Holothurians. But something unpredictable was happened.
(from left to right, in the photograph):
1. Marker for the length of DNA base pairs
2. DNA solution using sample A
3. DNA solution using sample B
4. PCR using sample A
5. PCR using sample B
6. PCR using sample C
7. PCR using sample D
Nb: Sample A was a similar species to sample B, but not for the individual. Sample A and B are of a different genus with either sample C or D. Sample C was a similar species with sample D, but not for the individual.
Hi- First, run your samples further next time, second, de-stain your gel longer or use less EBr - finally, I don't believe that your PCR worked (it looks like just primer bands to me). You should use controls and be sure that your PCR RX is working as well as trying different conditions and/or temps.
Best wishes!
-m.
I agree with Michael. Cant find find any target specific amplicons. Only primer dimers.
What a shit! I do not want to be rude. Just run a PCR with a negative control (a PCR with water instead of template DNA) and we'll be sure of what's going on. The PCR failed, of course. Regarding primer-dimers, we'd have a clue.
What is the expected size of the PCR band, and how would that relate to your marker? Without that information it is hard to give a lot of pointers, but it looks as if you did not get any amplification. The bands you are seeing might be primer dimers. I would re-do the PCR but include a positive and a negative control, and try to dilute your samples 1:10 or 1:20. You might actually have inhibition, and dilution your samples might help. Good luck!
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