I'm making a master mix by myself but there is no amplification with 16s or issr primers but it works with some specific primers I need a conclusion. Where is the mistake, and how can I solve it?
Sorry that it's not working for you. Let's start by seeing if we can narrow down the list of possible issues.
1. What does your positive control look like?
2. What size product are you expecting?
3. Does anyone else in the lab use these primers & have good results?
Let's start there and see if we can figure out what's going on.
thank you Katie for your response
1. I tried the same master mix with other specific primers for bacterial DNA and it gave the target band also with barcoding primers and issr primers and it worked also
2: It is supposed to give a 1000bp band
3. Yes, it works with other researchers in my lab
Katie A S Burnette
Sounds to me that the primers might have expired/degraded. Make a fresh dilution from the freezer stock and try again.
Good luck!
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