BIOT: various

[Arthur Bergen <bergen@AMC.UVA.NL>]

Note from the BIOT editors:

5 new BIOT messages, most of them technically oriented (RNA protocols,
DNA sequencing etc). Extensive protocols in response to these messages are
welcomed as messages to the entire HUM-MOLGEN BIOT TOPIC.
Please state your experience with these protocols.

Responses from commercial firms to HUM-MOLGEN BIOT are also welcomed, as
long as they pertain to ****alternatives methods techniques and methods
and clear assistance to the BIOT requests****.
We request commercial messages to be of clear assistance to our
subscribers, and not to much a clear add for the particular firm sending
the message. Messages with too much commercial slant(s) will be refused by
the editors without further notice.

Martin Kennedy
Arthur Bergen
***********************************************************************

From:   IN%"ED-MOLGEN@nic.SURFnet.nl"  "Human Molecular Genetics Editors" 20-OCT-1995 13:19:49.19
To:     IN%"ED-MOLGEN@nic.SURFnet.nl"  "Multiple recipients of list ED-MOLGEN"
CC:
Subj:   BIOT, CALL: New World Primate STR Primers wanted

Prof. Dr. H. Rothe from the University of Goettingen, Germany, is studying
the social organisation of Callithrichids. Now, he and his co-workers changed
the housing conditions of the Callithrix jacchus colony moving the animals
to a 20 acres semi free enclosure. Since extrapair copulations and/or
cooperative polyandry may occur tools for fatherhood analysis are necessary.
He will be much obliged to every DNA-specialist/geneticist for information
about pairs of STR (microsatellite) or VNTR primers specific for Callithrix
jacchus or other New World Primates. Please contact him at Institute fuer
Anthropologie, Ethologische Station, Sennickerode 11, D-37130 Gleichen,
Germany; phone: +49-5592-413, fax: +49-5592-1524 or e-mail to:
hzierdt@gwdg.de

Thanks, i.A. H. Zierdt
************************************************************************
From:   IN%"ED-MOLGEN@nic.SURFnet.nl"  "Human Molecular Genetics Editors" 24-OCT-1995 10:03:17.38
To:     IN%"ED-MOLGEN@nic.SURFnet.nl"  "Multiple recipients of list ED-MOLGEN"
CC:
Subj:   BIOT: Call for help for RNA Protocols

I would appreciate receiving information or protocols for obtaining total RNA
from fresh human EDTA blood samples. I have tried various kits such as RNAeasy
and RNAzol but have had little success. I wish to use the RNA for cDNA
synthesis and subsequent RT-PCR. Any helpful suggestions would be appreciated.
Yours truely,
Peter

**************************************************************************
From:   IN%"barzilay@icrf.icnet.uk"  "Gil Barzilay"  1-NOV-1995 15:04:34.69
Subj:   BIOT: Inducible expression systems

barzilay@icrf.icnet.uk (Gil Barzilay) sent the following comments:

------------------------------------------------------------
Dear all,

I would be very grateful if anyone has any hands-on experience using stratagene's
LacSwitch system or Gibco's Tetracycline inducible system.  I'm trying to
overexpress a DNA repair enzyme (HAP1) and although I can get it to work in
HeLa's in constiutive system, cloning it to the lacswitch or tet has resulted
in 0 induction/expression.  I've tried various conditions, all are transient
because I don't wish to grow stable colonies for months to realize that the
system works.

Any useful suggestions???

thanx, gil.

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Server protocol: HTTP/1.0
Remote host: mac026023.ox.icnet.uk
Remote IP address: 143.65.26.23

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From:   IN%"bzehnbau@imgate.wustl.edu"  "Barb Zehnbauer"  2-NOV-1995 22:12:57.32

bzehnbau@imgate.wustl.edu (Barb Zehnbauer) sent the following comments:

BIOT:
Can anyone recommend the optimal conditions for sending whole blood specimens for purposes of RT-PCR for a message >2.5 kb?
The RNA isolation kits we use is just fine for smaller RT-PCR products (< 500bp).  Is there some type of additive one can
add to the tube at the time the specimen is drawn which helps to stabilize the RNA?

Thanks for your suggestions,

Barb Zehnbauer, PhD
Washington University School of Medicine
Molecular Diagnostic Lab
314 362-1733
bzehnbau@imgate.wustl.edu

*****************************************************************
BIOT:
SUBJ: DNA sequencing

For reasons of comparision, I would be interested in some extensive
protocols for direct DNA sequencing of PCR products, pertaining to
mutation analysis of disease genes in humans. (including method of
cleaning up DNA, time request for protocol, perhaps a-symmetric PCR,
experience with that protocol, etc.)

Thanks,

Arthur Bergen
(Bergen@amc.uva.nl)