I did PCR analysis for my samples (total 6 samples), I got bands for the 5 samples, however, there is one sample which the band is detected at 10,000 bp? What should I do? Is there any relations with the volume of primers that I used?
I did 15ul total for each samples, and I used 5ul of DNA template for each samples. Any suggestions?
What is your product size?
Did you check con of your DNA in each samples?
10,000 bp???????
It is a plasmid?
I think it is your extraction DNA without any amplification.
I agree with Alireza Mordadi .
in this sample you may have added large amount of template DNA (>10Kb) this can sometimes inhibit the PCR reaction and in this case you will see no product and just your template DNA.
Hi!
As suggested, please give size(s) of the band(s), moles or nanograms of ingredients and upload the gel image. You should give the template amount (nano or micro gram), not in microlitre. So I think you're probably using too much template and this cab block PCR. For a 15 reaction I suggest adding the template in the range of 20-100 nanogram and this need to estimate the DNA concentration via spectrophotometer and dilute it prior to use.
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