lambda DNA prep

Martin Powell powellmb at cf.ac.uk
Mon Sep 11 06:48:40 EST 1995


I've been trying to isolate lambda phage DNA for 3 months, using liquid 
and plate lysates. I've had good results with the Stratagene Lambda DNA 
isolation kit. I grow phage on 140mm plates so that the whole plate is 
covered with plaques. I then elute at 4C with SM buffer for 24hrs. Then 
gently scrape the surface with a plastic loop to ensure all the phage 
comes off. Using the kit on 1ml of this eluant gives about 2micrograms. 
After finishing the kit, Stratagene were kind enough to tell me where the 
got all there reagents from, which means that you can effectively use 
your own reagent when some of theirs run out. I've found liquid lysates 
rather unreliable as I'm never sure if lysis has really occurred or my 
cells have died! At least with plate lysate you can detect if lysis has 
occurred. I've used this kit a number of times and I have found it to be 
more reliable than other methods I've used




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