Why is the primer I used for PCR exam is false , considering that the band was obtained in the first time but no band has been seen in the next series, how can i be sure about that the primer had a problem?
It appears you are new to molecular biology and facing problem just due to the lack of experience/expertise and knowledge. Sometimes, PCR or electrophoresis fails just due to the carelessness of user. I recommend you to get help from someone more experienced in your lab in PCR assays.
By the way, your question does not say enough to be answered.
Hi Mohammad Karbakhsh Ravari , I agree with Abhijeet Singh that your question is not so clear. For a full answer you need to give us more details. Which size is the band do you expect? did you check that your primers are specific for your target? do you have a positive control?
Do you have some of the original sample to re use as a positive control sample?
What does your gel look like.....is it completely clean or is there a smear in the sample track. Is there a diffuse small band in your sample tracks? Can we see a picture of the gel image?. Was there a very long time between the working experiment and the failed pcr?
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