How to make high titre lambda stock?

Csaba Jeney jencsa at xenia.sote.hu
Mon Dec 7 13:24:52 EST 1998


On 6 Dec 1998 19:52:19 GMT, "Allison B. MacKay"
<mackayab at biology.queensu.ca> wrote:

>I am trying to make a high-titre stock of phage lambda (EMBL3)
>in order to generate large working quantities of my genomic
>clone.  I have tried plate and liquid lysis protocols from
>several sources (mainly Sambrook and Short Protocols) but
>only seem to get maximum titres of 10e7 to 10e8 pfu/ml.  The
>protocols strongly recommend using stocks of 10e9 to 10e10
>pfu/ml to infect larger cultures.  Is there some trick that
>I don't know about that will boost the yield?
>
>Allison Mackay
>Department of Biology
>Queen's University
>Kingston, Ontario

We also had this problem, but finally it is solved, use plate lysis
intsead of liquid lysis. In our hand the different liquid lysis
protocols resulted in a low titer (10e8 used to be, but telling the
truth only once we got 2*10e10, but we could not repeat it anymore).
But after some optimisation our plate lysis protocol could result
repeatedly a 5*10e9 pfu/ml titer. Send me an e-mail for details.

Good luck

Csaba Jeney



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