LIBRARY PLATING: no lysis of bacterial lawn

Kevin O Hwang khwang at SPECIALK.IAIMS.BCM.TMC.EDU
Wed Apr 6 17:47:19 EST 1994


HELP!
We are attempting to plate out two libraries, one is a lambda ZAP and
the other is lambda gt10. The host cells for the ZAP library are from
an LB plate and are XL1 blues. The gt10 host cells are C600 hfl, and
are from a frozen stock, streaked out on a plate and a single colony
chosen. The cells are both grown overnight in 10 mM MgSO4 and 0.2% maltose.
They are spun down, resuspended in 10 mM MgSO4 and used at an OD of .5
The libraries are diluted in SM and plated out on fresh, dry NZY plates
following incubation of the bacteria and phage for 15 min at 37C. The
resulting lawn is PERFECT but, for the third time, there is NO LYSIS,
no clearing, nothing!!!! Does anyone have any ideas? I've screened over
20 million phage previously, and the library (at least one of them)
is known to be good (as of 6 months ago, and it's been stored at 4C since
that time).

Many thanks in advance
Kevin and Jill



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