Help with Lambda DNA isolation
Jean L Dowty
jldowty at unity.ncsu.edu
Wed Dec 7 16:47:25 EST 1994
RiverSloth (riversloth at aol.com) wrote:
: In article <pabkwc.2.000F4A21 at leeds.ac.uk>, pabkwc at leeds.ac.uk (K.W.
: Cheung) writes:
: >Can someone recommend a good method for Lamda DNA isolation. >I've tried
: using
: >the Qiagen midi prep using plate lysate but only got small amount >of DNA
: back.
: > What is the best host for the propagation of Lamda DASH II ?
: >
: I have had the best success using Lambda Sorb. I've tried 4 or 5 methods
: from Maniatis and the Red Book and other sources and found LambdaSorb to
: yield much better amounts and gives DNA which can be restriction digested.
: Other methods I have tried do not yield much DNA which is clean enough to
: cut with enzymes.
: Be sure and follow the product instructions and use agarose plates and not
: agar plates.
: Good luck
: Felix D. Guerrero
: USDA-ARS
: Kerrville, TX
I have also had no luck with the Quiagen kit and someone suggested that
you need to start out with AT LEAST 2x10(10) pfu for any kind of yield
(1ug). Most people I have talked to about lambda have protocols using
LE392 as the host.
Just my two cents
JL
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