Lamda phage - RIP?

martin LEACH leach at bu.edu
Thu Feb 13 20:08:02 EST 1997


David L. Haviland, Ph.D. (dhavilan at IMM2.IMM.UTH.TMC.EDU) wrote:
: Hi all:

:  I have a very dear friend who ordered the construction of a lamda phage
: cDNA library from a retputable company.   It was made to "specs" and showed
: promise.  It unfortunately arrived when he was out of town and the
: "brain-dead" idiot that received the box, in my collaborator's abscence,
: simply put the whole thing (containing the phage stock) at -20'C.  The
: company did not add glycerol or DMSO to the phage stock prior to shipment.

doh!

1. titer the phage and see what the damage really is.

: Akin to plasmid resuce from dead cultures of bacteria, is anyone (Martin,
: Jim, Paul?) aware of any method to salvage the DNA from the phage stock -
: which now likely has the big *zero* for a titer instead of 6X10^6 that it
: orginally was.  Any way the DNA might  likely be extracted (reliably) and
: re-packaged?

2. I'd try a standard phage prep...(isolate the DNA) and try repackaging
it...will probably get some shearing and loose the real big stuff......if
really worried about big stuff....try solid-phase extraction similar to how
DNA is prepp'd for PFGE. (modified for phage instead of yeast etc..)
....I'm guessing here...

: We are in the process of trying to get more of the library from the company.

3. lie....tell them they sent it to the wrong place and you just got it ;)
...never tried this...usually don't work though! - could even try blaming it
on fedex etc... for letting the sample freeze during shipment.....(we were
having some reagent problems from ABI/PE once...it appears that the capping
reagent for the synthesizer was being frozen while shipped...and was arriving
with crystals.......perhaps you could use this angle.........my nefarious side
is coming out tonight ;)

: Is it the big DIA (do it again)?


: ::sigh::

time to kick that tech up the dewberries!

M


--
.....          Martin Leach                Email:leach at bu.edu
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