DNA extraction - cell treatment for gram positive cocci (Sep/27/2006 )
do you know any protocol for Staphylococcus aureus cell lysis treatment before DNA extraction using achromopeptidase? it usually used in PFGE but i want to use it for PCR. i used lysostaphin before but i ran out of it so i just wondering if achromopeptidase can be use for this. thanks a lot!
Hi icemizu, I am also working with Staph.aureus cells but I use lysostaphin to break cell wall though I have heard of using achromopeptidase in one of the papers.I will certainly try to find out about it for you.
I work on strep
3-4 colonies in 100 microliter of 50mMNaoH 95 degrees water bath for 1 min
and then cool to 4 degrees
Then add 16 microliter of 1M Tris HCl pH8
centrifuge at high speed for 2 mins..template in the supernatant...
shoud work fine
thanks bela n tigress. i did find one method in paper but i juz wanna know if there is other method for it. i loved to share it with u. looking foward for the next reply!!
p/s: this is the paper i found.
"Modified DNA extraction for rapid PCR detection of MRSA", Iranian Biomedical Journal 8 (3): 161-165 (2004)