Fw: Cosmid library screening

Malay curiouser at ccmb.ap.nic.in
Sun Sep 19 04:40:11 EST 1999


This is my third posting hoping that someone will reply

Malay

----- Original Message -----
From: <curiouser ("Malay")>
To: <methods at hgmp.mrc.ac.uk>
Sent: Saturday, September 11, 1999 11:08 AM
Subject: Fw: Cosmid library screening


: This is my second posting
:
: Malay
:
: ----- Original Message -----
: From: <curiouser ("Malay")>
: To: <methods at hgmp.mrc.ac.uk>
: Sent: Wednesday, September 08, 1999 3:14 PM
: Subject: Cosmid library screening
:
:
: > Dear Netters:
: >
: > I've made a cosmid (pLAFR3) genomic library of Pseudomonas. My library
is
: > in 96-well titer plates and I generally grow the colonies on agar
plates
: > overnight on membranes. I use hybond-N+. I am screening the library
with
: 5
: > different probes. I have a plan to screen with all the probes at one
: time,
: > i.e. to add all the labeled probes at one time in the hybridization
: buffer.
: > I am using Church's buffer (0.5M Na-phosphate, 7%SDS) for
hybridization.
: > But the signal to noise ratio is very poor. I can hardly make out any
: > difference. Can anyone help me out?
: >
: > I am particularly looking for the information regarding any special
: tricks
: > of colony blots, hybridization, how much count I should add to buffer
: etc.
: >
: > Please help me out.
: >
: > Malay
: >
: > Malay Kumar Basu
: > Centre for Cellular and Molecular Biology
: > Hyderabad 500007
: > I N D I A
: >
: > Fax: (00-91)40-7171195
: > Phone: (00-91)40-7172241
: > -----
: > Alone: In bad company.
: > -----
: > curiouser at ccmb.ap.nic.in
: >
: >
: >
: > ---
:
: ---

---



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