Identification of Listeria monocytogenes - (Aug/18/2014 )

Hello to all,

I have isolated Listeria monocytogenes from my food samples and I would like to identify the strains by PCR method.

your help is appreciated in these 3 points (if possible please):

1-I havent done this before or worked with L.Mono, but i have been reading that targeting the hly gene would identify the strains that i have. What other genes that could also be specific for L.Mono that i should target?

2- has anyone worked with the L.Mono identification by PCR and can share with me the protocol?? I need to order all reagents and primers that are essential to run this work. would be great when I get an advice.

3- My Listeria monocytogenes isolates are preserved in 30% glycerol (freezed). For DNA extraction,  Should I directly streak a loopful from the glycerol preserved stock onto the agar plate or I should better first dispense it in TSB for overnight at 37C then later streak it ?

I am not sure what is the best way to recover my Listeria glycerol stock and optimum subculturing methods to maintain my bacteria !

I hope to hear from you!

thanks in advance,

DIma

 i just found an answer on my earlier posts concerning the detection of virulent genes in Listeria mono.:

the "microseq listeria monocytogenes "detection kit..