I'm looking for best parameters for the PCR as well as best primers which I can use while investigating lactic acid bacteria. I was thinking of using M13 as a primer sequence, but I'd be glad if anyone could give me some advice. Thank you very much in advance.
you can also use the ITS regions for that purpose
the sequences for different ITS regions can be googled........PCR condition will change according to primer pairs annealing teperature..........one have to standardise it roughly the annnealing temperature is 3 degree lesser than the lowest melting temperature of primer pair
Hello Szymon,
Rep-PCR using the (GTG)5 primer has been proven to give very good results. It has been tested in Lactobacilli and Enterococcus and creates fingerprints which allow differentiation to the species level. Have a look at the attached files but there's a lot more of them using (GTG)5. You may have to do some optimization with regards to magnesium concentration and annealing temperature in your PCR in order to get satisfying resolution and clarity, but I believe that the result is very satisfying.
All the best
Panos
Hi Szymon: we had good results with P32 primers. Please check below the paper by Cusick and Sullivan (2000) in Appl. Environ. Microbiol.
http://aem.asm.org/content/66/5/2227.full
Best regards
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