Small scale lambda growth

Michael R. Leonardo mrl at csd.uwm.edu
Sat Nov 25 15:23:28 EST 1995


Sorry all, my server tends to eliminate the post.
Here is a protocol for a small scale Lambda isolation...again

Grow your host strain in 5ml tryptone broth (TB) overnight at 37C
Place the culture in an Oakridge tube and spin for 10 min at 1500 x g
Discard supernatant and resuspend the pellet in 2.5ml 10mM MgSO4
Place 50ul in 18mm culture tube
Add either 20ul lysate or 1-2 plaques
Vortex 5 seconds and let stand 5 min at RT
Add 2ml TB + 10mM MgSO4
Incubate 4-6 hours at 37C (til lysis)
Add 100ul CHCl3 and vortex
Transfer lysate to either Oakridge tube or eppi (1.5ml)
Spin 10 min at 4500 x g
Transfer S/N to fresh screwcap or eppi
Add 100ul fresh CHCl3 and store at 4C
Titer as normal

This has worked well for me over the years.  I hope this helps.

Michael Leonardo, Ph.D.
Dept. Biological Sciences
UW-Milwaukee
P.O. Box 413
Milwaukee, WI 53201

(414) 229-6373
mrl at csd.uwn.edu





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