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lambda expression screening

Dave Knecht KNECHT at UCONNVM.UCONN.EDU
Wed Aug 18 12:05:33 EST 1993


Hi netters- I have a question regarding the growth of lambda for antibody expre
ssion screening.  Sambrook et al. has a protocol for growth of phage libraries
in which they grow bacteria in maltose supplemented media, infect and then plat
e at 37C.  In a separate section for antibody screening, they infect and then g
row at 42C for several hours before switching to 37 for IPTG induction and filt
er overlay.  I can see no obvious reason why the 42C step should be needed.  Th
e only ts gene I know of in the lambda is the repressor which should only affec
t lysis vs. lysogeny.  IN fact, I presume 42C is not necessary for lysis under
the conditions used ( and in our hands so far is not).  Does anyone out there u
nderstand why Sambrookindicates 42 should be necessary for screening?  Also, wh
en we switched from a gt11 library in Y1090 to a Zap library in XL1 blue we had
 trouble with growth of phage, bacteria or both.  Is there some alteration to t
he growth protocol necessary from others experience?

Dave Knecht
Knecht at unconnvm.uconn.edu



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