LLNL HUMAN SINGLE CHROMOSOME COSMID LIBRARY :
Chromosome Y
LL0YNC03
"M"
About the library
This Chromosome Y cosmid library was constructed at LLNL. It has an
average insert size of between 32-47 Kb and contains 12,480 clones providing
a 7-fold coverage of the chromosome. The vector used is Lawrist 16. The
library consists of 33 plates labelled 1-33.
Production of high-density gridded filters
The library has been gridded in a 4x4 array on one 22
x 22 cm Hybond N nylon membrane (Amersham). Each clone has been spotted
twice to give 24,960 clones (12,480 x 2) on the membrane which is labelled
"M".
Clones spotted on the filters were grown overnight at 37°C
on LB agar plates containing Kanamycin (20µg/ml). The filters were
subsequently processed by (a) SDS treatment, (b) denaturation and (c)
neutralisation, then dried and crosslinked using a Spectronics Corporation
XL-1500 UV Crosslinker. The filters are ready for pre-hybridisation in
appropriate buffers.
User Information:
For successful removal of probes, and to enable re-probing, membranes
must NEVER be allowed to dry during, or after, hybridisation and washing.
Membranes can be stripped to facilitate re-probing by washing sequentially
at RT in 0.2M NaOH for 30 min. and 0.1 x SSC / 0.1% SDS / 0.2M Tris-HCl pH 7.5 for 15 min. A gentler method, which extends the number of probings
per filter, is to wash the probed membrane after use in a large volume
of pre-hybridisation buffer overnight at 65°C, prior to using
another probe.
Subsequent publication:
Please acknowledge both the originator of the library, LLNL as described
in the "materials transfer agreement" that you have signed,
and Geneservice for providing the library, in any publication
arising out of using this resource. An example of how a clone (E.g. CLONE
NAME = 32-B9) should be described in a publication is: M32b9 from the
LL0YNC03 library.
5th Sept 2000 Version 1.2
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