Question on Replica plating

CVRL microbio at emirates.net.ae
Mon Apr 20 03:43:08 EST 1998


I am interested in screening a large number of bacterial colonies for
microsatellites. Can someone suggest a simple method for transferring
colonies to nitrocellulose filter paper, apart from using a replica
plating tool and velvet cloth, Sambrook et al.  Would laying the filter
over the colonies for a while result in sufficient binding of bacteria
for subsequent lysis and fixation? 

My other question concerns a suitable method for binding DNA to
nitrocellulose filters. Most manuals mention baking at 80 degrees for
1-2 hrs in a vacuum oven. Now, we don't have a vacuum oven, so is there
a way to get around this? Baking longer? 

Thanks in advance for all your assistance. Please direct your response
to my  e-mail address, as I don't always have access to this newsgroup. 

Regards,
Maxy

Maxy Mariasegaram
Central Veterinary Research Laboratory
P.O. Box 597
Dubai
United Arab Emirates
							
E-mail: microbio at emirates.net.ae



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