HDTenEightyP/NetNews huggingface dataset:
From: excoffie@sc2a.unige.ch (Laurent Excoffier)
Newsgroups: bionet.biology.computational
Subject: Analysis of Molecular Variance : A Windows 3.0 program
Date: 8 Sep 92 13:13:52 GMT
Distribution: bionet
Originator: biocomp@lerkim.umiacs.umd.edu


  This is an annoucement for the public release of a WINDOWS 3.x based program

to analyze the GENETIC STRUCTURE OF POPULATIONS using MOLECULAR INFORMATION.


  This program (WINAMOVA ver 1.04) is based on a paper by Excoffier, Smouse and 

Quattro (1992), and computes variance components for 2 levels of population 

structure as well as their associated modified F-statistics (called here Phi-

statistics).


  This version can handle a maximum of 255 molecular haplotypes, 100 populations

and 100 groups of populations. It requires haplotype frequency information for

each population, as well as a matrix of inter-haplotypic squared (euclidian) 

distances. 


  Conventional multi-allelic F-statistics may also be computed.


  The program includes a non-parametrical (permutational) test for the 

significance of both variance components and Phi-statistics.


  It can be uploaded by anonymous FTP from    : ACASUN1.UNIGE.CH

(username : anonymous) in the directory       :  pub/amova

under the form of an archived file called WINAMOVA.ZIP, including 

executable versions for Windows 3.0 and Windows 3.1, plus example files.


           The program is FREEWARE and can be distributed freely in its 
           original form, but I would appreciate receiving an E-mail from
           interested users, as well as comments, suggestions, and bug
           reports.

(Windows 3.x is a product from Microsoft Corporation)


  Reference cited :
  =================

Excoffier, L, Smouse, PE, and Quattro, JM (1992) Analysis of molecular variance
      inferred from metric distances among DNA haplotypes: Application to human
      mitochondrial DNA restriction data. Genetics 131:479-491.


*******************************************************************************
*                                                                             *
*  Laurent Excoffier                					      *	
*  Genetics and Biometry Laboratory           Tel    : 41-22-343.69.30        *
*  Dept. of Anthropology and Ecology 					      *	
*  University of Geneva                       Fax    : 41-22-300.03.51        *
*  12, rue Gustave Revilliod                  				      *	
*  1227 Carouge-Geneva, Switzerland           E-mail : excoffie@sc2a.unige.ch *
*									      *	
*******************************************************************************
From: sashazh@face.msk.su
Newsgroups: bionet.journals.contents
Subject: Biological Membranes (Moscow). 1992. v.9, No.2.
Date: 27 Aug 92 01:04:06 GMT
Distribution: bionet


CC Journal: Biological Membranes (Moscow). 1992. v.9, No.2.

AU Muzykantov-V-R.  Puchnina-Artushenko-E-A.  Chekneva-E-V.
   Voyno-Yasenetskaya-T-A.
TI Activation of the phosphoinositide turnover in human endothelial  cells  by
   sublethal doses of hydrogen peroxide.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 133-142.

AU Molotkovskaya-I-M.   Svirshchevskaya-E-V.   Litvinov-I-C.    Mikhailov-I-I.
   Dyatlovitskaya-E-V.  Molotkovsky-Jul-G.  Bergelson-L-D.
TI Immunosuppressive  activity  of  glycosphingolipids:   a   study   of   the
   interleukin-2 with gangliosides using cells and model systems.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 143-151.

AU Marchenko-S-M.  Sagach-V-F.
TI Effect  of  methylene  blue  on  the kinetics of Ca(2+)-activated potassium
   channels.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 152-157.

AU Gukovskaya-A-S.  Kotelevskaya-S-M.  Trepakova-E-S.  Zinchenko-V-P.
TI Effect of SH-reagent thimerosal on Ca(2+) homeostasis in thymocytes.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 158-165.

AU Marinov-B-S.  Filatov-G-N.  Niconov  S-S.  Ruzhieva-R-Ch.  Lubarsky-A-L.
   Fesenko-E-E.
TI Regulation of conductance of retinal rod outer segment membrane by redox
   agents.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 166-173.

AU Marinov-B-S.  Philippov-A-K.
TI Relevance of the redox properties of some Ca channel modulators to their
   action.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 174-183.

AU Dubovski-P-V.  Esipova-O-V.  Vasilenko-I-A.  Zvonkova-E-N.
TI A wide-line NMR study of hydrophobic peptides and their behaviour in the
   phospholipid bilayer.  III.  Comparision of Gly-Leu-Phe peptide with its
   N-acylated, O-alkylated derivatives.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 184-192.

AU Gordeliy-V-I.  Islamov-A-Kh.  Syrykh-A-G.
TI Determination  of  repeat  distance fluctuations of lipid and biological
   membranes via time-of-flight neutron diffraction.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 193-200.

AU Ermakov-Yu-A. Cherny-V-V.  Sokolov-V-S.
TI Adsorption of beryllium on neutral and charged lipid membranes.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 201-213.

AU Markin-V-S.  Isong-T-Y.  Robertson-B.  Astumian-R-D.
TI Membrane transporters as free energy transducers: optimisation of energy
   transfer.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 214-222.

AU Pastushenko-V-Ph.  Chizmadzhev-Yu-A.
TI Energetic  estimations  of  the deformation of translocated DNA and cell
   membrane in the course of electrotransformation.
SO Biol-Membranes-(Biologitcheskie-Membrany).  1992 Mar.  9(2).  P 223-232.
Newsgroups: bionet.journals.contents
From: davidg@ccsg.tau.ac.il (gutnik david)
Subject: Bio-Journals
Original-To: Bio-Journals@uk.ac.daresbury
Distribution: bionet
Date: Fri, 11 Sep 1992 05:51:38 GMT
X-Mailer: ELM [version 2.4dev PL52]

I would like to renew my subscription to Bio-Journals. Please note my
new e-mail address: davidg@ccsg.tau.ac.il
Thanks,
David Gutnick
From: kristoff@genbank.bio.net (David Kristofferson)
Newsgroups: bionet.journals.contents
Subject: Russian Language Journal TOCs
Date: 8 Sep 92 17:30:58 GMT
Organization: GenBank Online Service

Note to our readers:

Alexander Zhuikov has volunteered to post translations of Russian
language journal TOCs to help promote greater awareness of biological
research originating in his country.  I hope that you find these
postings to be useful.

				Sincerely,

				Dave Kristofferson
				BIOSCI/bionet Manager

				kristoff@genbank.bio.net

Newsgroups: bionet.journals.contents,bit.listserv.bionews,sci.bio
From: bsp1sz@surrey.ac.uk (ZHANG  S  MR)
Newsgroups: bionet.journals.contents
Subject: subscription
Date: 9 Sep 92 21:33:52 GMT
Distribution: bionet
Mailer: Elm [revision: 66.25]
Original-To: bio-jrnl@uk.ac.daresbury

I would like to subscribe Bio-Journals please.
Newsgroups: bionet.journals.contents,bit.listserv.bionews,sci.bio
From: fredh@matt.ksu.ksu.edu (Fred Hunt)
Newsgroups: bionet.journals.contents
Subject: Wanted: Info on net bio journals
Date: 11 Sep 1992 18:08:25 -0500
Organization: Kansas State University
NNTP-Posting-Host: matt.ksu.ksu.edu

Hello,
	I am new to the network and I am interested
in journals and newsletters available through email, 
Internet, and BITNET. If any one has any suggestions
please email them to me. Thank you for your time and
effort, I really appreciate it.
-------->
Fred Hunt
fredh@matt.ksu.ksu.edu
From: djt2@po.CWRU.Edu (Dennis J. Templeton)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: screening phagemid libraries
Date: 11 Sep 92 17:41:34 GMT
Reply-To: djt2@po.CWRU.Edu (Dennis J. Templeton)
Organization: Case Western Reserve University, Cleveland, OH (USA)
NNTP-Posting-Host: thor.ins.cwru.edu


In a previous article, horton@molbio (Robert Horton) says:

>Greetings Netlanders!
>
>	I have been using replica plating and colony hybridization to 
>screen "micro-libraries" in a plasmid. This seems to be practical for small
>numbers of colonies, but if I decide to try screening over about 1000, I
>want a better way. First I thought I'd make a mini-library in m13, which
>I can screen by plaque lifting. However, this would require that I re-clone
>the insert into my plasmid once I've identified it. Here's my question:
>since my plasmid (pBluescript II) has a single-stranded phage origin (it is
>a "phagemid"), is there some way to screen them as plaques? I figure if the
>helper phage doesn't lyse the bacteria very well all by itself, one might
>be able to get plaques only where the Bluescript clones are...?


We've been using libraries that we shift from lambda form (ZapII) to
phagemid, to ssDNA, and are worried about retaining complexity from one
step to the next.  Frankly, the step I'm most worried about is making the
ssDNA from the phagemid; if some clones grow much faster they will swamp
out the slowgrowers and we will lose complexity.

If we were doing something as simple as colony/plaque lifts, I'd be
satisfied to keep the library in plasmid form, and screen ampR colonies...
that way you are more assured that complexity is retained.

Just one man's opinion...

dennis
Newsgroups: bionet.molbio.methds-reagnts
From: jjw@rsbs0.anu.edu.au ( Jeremy Weinman)
Subject: dye-term removal after cycle seq rxns
Organization: Research School of Biological Sciences, Australian National University
Date: 10 Sep 92 09:52:29 +1000

Hello fellow netters,
	I've a question regarding clean-up of dye-terminator cycle sequencing
reactions.  Specificaly, does anybody out there have any nifty, simple, 
cheap protocols for using home-made spun columns for this step?  I've used
G-50 packed Eppies spun in a Speed-Vac before to remove unicoprorated counts
after probe making, but I'm uncertain if this method is going to remove 
sufficient dye-terminators to give good clean sequence.  I'd rather not 
waste reactions to find out if somebody already knows the answer.
	Any helpful suggestions would be most welcome,
		Cheers,
			Jeremy
=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
Dr. Jeremy Weinman				Email:	jjw@rsbs1.anu.edu.au	
Plant Microbe Interaction Group			Phone:	61 6 2495051
Research School of Biological Sciences		Fax:	61 6 2490754
Australian National University			Snail:	PO Box 475, Canberra, 
							ACT 2601, AUSTRALIA
From: jing@milton.u.washington.edu (Jing Huang)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Help: Transfection into mammalian cells
Date: 9 Sep 92 01:35:34 GMT
Reply-To: jing@u.washington.edu
Organization: University of Washington, Seattle

Hi, I am trying to co-transfect 2 plasmids into both 3T3 (fibroblasts) 
and C2 (myoblasts) cells.  I am doing transient transfection and I need
to be able to control both the ratio of the 2 plasmids and the absolute
copy number of each plasmid inside a single cell.  The ratio can be 
achieved by the calcium phosphate method which gets multiple copies of 
plasmids into a cell, while electroporation usually gets only 1 copy of 
plasmid into each cell.  Does anyone know how to limit the copies of 
DNA molecules in the Ca method?  and how to get more than 1 but defined 
number of plasmids by electroporation?  Thanks! and I will post a summary.

-Jing
jing@u.washington.edu
From: jak8@po.CWRU.Edu (Joel A. Kreps)
Newsgroups: bionet.molbio.methds-reagnts
Subject: long-ranger gels?
Date: 15 Sep 1992 14:45:05 GMT
Organization: Case Western Reserve University, Cleveland, Ohio (USA)
NNTP-Posting-Host: slc5.ins.cwru.edu


Dear Network,

I will be doing quite a bit of sequencing soon in search of a 
mutation and I would like to hear from people who have used the
long-ranger product from AT biochem.  Specifically, how much 
better is it when compared to normal acrylamide, are there any
problems with pouring long gels (ie 80 cm long)?  Any tips would
be much appreciated.

Joel A. Kreps
Case Western Reserve Univ.
Newsgroups: bionet.molbio.methds-reagnts
From: horton@molbio.cbs.umn.edu (Robert Horton)
Subject: Re: mRNA from bovine PBMC
Organization: University of Minnesota
X-Newsreader: Tin 1.1 PL5
Date: Mon, 14 Sep 1992 17:57:35 GMT

KENNEDY, DEBBIE A (dak7826@zeus.tamu.edu) wrote:
: 
: 1 x 10^9 cells is yeilding ~100 ug total RNA and 25 ug mRNA.
:                             ^^^^^^^^^^^^^^^^^
How much total RNA?
--
Bob Horton   (horton@molbio.cbs.umn.edu)
Newsgroups: bionet.molbio.methds-reagnts
From: horton@molbio (Robert Horton)
Subject: screening phagemid libraries
Organization: University of Minnesota
Date: Thu, 10 Sep 1992 17:43:52 GMT

Greetings Netlanders!

	I have been using replica plating and colony hybridization to 
screen "micro-libraries" in a plasmid. This seems to be practical for small
numbers of colonies, but if I decide to try screening over about 1000, I
want a better way. First I thought I'd make a mini-library in m13, which
I can screen by plaque lifting. However, this would require that I re-clone
the insert into my plasmid once I've identified it. Here's my question:
since my plasmid (pBluescript II) has a single-stranded phage origin (it is
a "phagemid"), is there some way to screen them as plaques? I figure if the
helper phage doesn't lyse the bacteria very well all by itself, one might
be able to get plaques only where the Bluescript clones are...?
	Any comments or suggestions would be appreciated, and if we 
figure anything out, I'll post it.

--
Bob Horton            /\ "Crash programs fail because of the theory that
U. of Minnesota, CBS  || with nine women pregnant you get a baby a month" 
1479 Gortner Ave.    /||\   -Werner von Braun.  Disclaimer:"Bob who?"
St. Paul, MN 55108    ^^   horton@mobio.cbs.umn.edu/(612) 624-3790
Newsgroups: bionet.molbio.methds-reagnts
From: dak7826@zeus.tamu.edu (KENNEDY, DEBBIE A)
Subject: mRNA from bovine PBMC
News-Software: VAX/VMS VNEWS 1.41    
Organization: Texas A&M University, Academic Computing Services
Date: Fri, 11 Sep 1992 21:18:00 GMT


	Am currently isolating total and messenger RNA from mitogen
stimulated bovine lymphocytes, yield is low, purity excellent.....
1 x 10^9 cells is yeilding ~100 ug total RNA and 25 ug mRNA.Bovine
spleen yields much more,.....is it me or the cell source, any info, comments,
snide remarks are welcomed!!!!!!

Thanks in advance,
d.
Newsgroups: bionet.molbio.methds-reagnts
From: bchs1b@Rosie.UH.EDU (Michael Benedik)
Subject: Re: Lambda EMBL
Reply-To: bchs1b@Rosie.UH.EDU
Organization: University of Houston
Distribution: bionet
Date: Tue, 15 Sep 1992 05:30:13 GMT

In article <1992Sep15.023903.16475@news.iastate.edu>, gat44@iastate.edu (Gabriel Trueba) writes:
>Dear Netters
>
>Does anybody know why E.coli strain 1090r-(RecA+) is not 
>used for lambda EMBL libraries?. I am aware of the fact
>that P2 lysogens are the best.
>
>gat44@iastate.edu
>
>

I don't know the genotype of 1090r- off the top of my head, but there
probably is no specific reason why it isn't used. However, for reasons
not always known, lambda forms nicer plaques and higher titers on some
hosts than it does on others. Therefore the hosts you see recommended 
for use with lambda are usually those which, in peoples experience, are
known to work well. This does not mean that other hosts won't work.

You only need to use a P2 lysogen for the initial plating/amplification
of your library. Afterwards you don't need to continue using P2 lysogens.

----------------------------------------------------------------------
 Michael Benedik				INTERNET: Benedik@uh.edu
 Dept. of Biochemical & Biophysical Sciences	
 University of Houston				BITNET: Benedik@uhou
-----------------------------------------------------------------------
From: mic236u@vaxc.cc.monash.edu.au
Newsgroups: bionet.molbio.methds-reagnts
Subject: Frozen competent RecA- strains
Date: 7 Sep 92 12:45:36 +1000
Organization: Computer Centre, Monash University, Australia

Hi Fellownetters,

Some time ago I read somewhere that competent RecA- cells don't like to be
frozen; they lose their competence in a couple of weeks. At the time it 
explained the problems I had with my frozen competent cells. (Unfortunately, 
I did not keep the reference.) 

When I came in this lab I found that here every one or two months a fresh 
batch of competent cells (DH5alpha) is made and stored at -70C. This works 
fine most of the time. However, recently we had problems getting 
transformants from ligations whereas cccDNA worked fine. 
I hope a fresh batch of competent cells will solve our problem.

But some questions are inevitable:

- Has anyone out there the reference I forgot to keep?

- Can cells lose competence for oc/ccDNA but stay competent for cccDNA?
  I know: transformation efficiency cccDNA >>> oc/ccDNA. But we got good 
  numbers of colonies with cccDNA and zero with several ligations, and
  the ligations worked! What is the difference in the uptake of cccDNA 
  and oc/ccDNA; is it just a matter of size? 

- Has anyone a good method to keep RecA- strains competent for a long 
  time at -70C? Some companies sell frozen competent RecA- cells. For 
  how long are they guaranteed to work? 

- Do other Rec- strains (RecF, B, C) also loose their competence in the 
  freezer?

Many thanks in advance

Cheers Volker

PS  cccDNA : supercoiled DNA
     ocDNA : open circular DNA (ligation with phosphatased vector)
     ccDNA : closed circular DNA  
Newsgroups: bionet.molbio.methds-reagnts
From: fulton@binah.cc.brandeis.edu
Subject: Re: Tacking down cells
Reply-To: fulton@binah.cc.brandeis.edu
Organization: Brandeis University
Date: Fri, 4 Sep 1992 21:44:47 GMT

I believe Collaborative Research took over Cell Tak.
From: pnh@fcsparc6.ncifcrf.gov (Paul N Hengen)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: screening phagemid libraries
Date: 11 Sep 92 15:38:39 GMT
Organization: Frederick Cancer Research and Development Center

In article <1992Sep10.174352.27327@news2.cis.umn.edu> horton@molbio (Robert Horton) writes:

|>Greetings Netlanders!
|>
|>I have been using replica plating and colony hybridization to 
|>screen "micro-libraries" in a plasmid. This seems to be practical for small
|>numbers of colonies, but if I decide to try screening over about 1000, I
|>want a better way....[edited]

|>... Here's my question:
|>since my plasmid (pBluescript II) has a single-stranded phage origin (it is
|>a "phagemid"), is there some way to screen them as plaques? I figure if the
|>helper phage doesn't lyse the bacteria very well all by itself, one might
                       ^^^^
|>be able to get plaques only where the Bluescript clones are...?

|>Bob Horton

Bob: The life cycle of M13 does not include lysing of the host cell.
The infected cells grow slower than uninfected cells giving a turbid
plaque. Perhaps you could simply infect the entire library population
with a wildtype M13 and hybridize the lifted plaques.

Paul N. Hengen
National Cancer Institute
Frederick Cancer Research and Development Center
Frederick, Maryland 21702-1201 USA
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: lambda won't cut
From: cytogen@chmeds.ac.nz (Martin Kennedy)
Date: 10 Sep 92 11:11:55 +1300
Distribution: bionet


In article <9209091448.AA23264@server.uga.edu>, mcgraw%gandal.dnet@SERVER.UGA.EDU writes:
> 
> 
> Hello netters,
> 
> A colleague provided us with some lambda clone DNAs "purified" from plate
> lysates.  They won't cut with EcoR1.  We tried further purification with
> additional phenol/chloroform extractions.  The uncut DNA looks good on a
> gel, and the OD's are good.  The inserts should cut out with EcoR1 based on
> the way the library was made (R1 linkers).  We haven't worked with lambda
> in quite a while, but I imagine that some of you out there are cutting
> yours successfully as you read this.
> 
> What do you suggest?
> 
>                                 Mike Grosse
>                                 McGraw lab
>                                 University of Georgia
> 




Mike - 

Have you tried adding spermidine to 4mM in your digetions? This can make  a big
difference with lambda preps.  Also, try cutting less DNA in a bigger volume.

Cheers, 

Martin
From: bsh@MED.PITT.EDU (Basavaraju Shankarappa)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: AT Biochem - address etc??
Date: 10 Sep 92 15:06:22 GMT
Distribution: bionet

> Hello M&M's..
> 
> Recently, lurking in the back of Nature, I found a mention of
> a product called Hydrolink MDE (mutation detection enhancement)
> gel. It is purportedly capable of resolving heteroduplex fragments
> with 1 bp change in 900.
> 
> However, Nature failed to give the address, phone, fax, email (hah hah)
> of this company - I suspect they're on t'other side of the pond.
> 
> Can any of you help. If anyone has any experience of this product, I'd
> like to hear from you too.
> 
> Rob Beynon, Biochemistry, Liverpool
> 

J.T. BAKER now sells hydrolink products.  

The toll free number in US is 1-800-JTBAKER (1-800-582-2537).  I am sure
they must have a subsidiary or something of that nature in your part of th
world.
Raj Shankarappa
Univ of Pittsburgh
FR*ATLAS*CICT; arrival Fri, 11 Sep 92 16: 49:51 +0100 action Relayed
From: CALCIUM@FRCICT81.EARN
Newsgroups: bionet.molbio.methds-reagnts
Subject: phage sequencing
Date: 11 Sep 92 15:49:51 GMT
Distribution: bionet
P1-Message-Id: FR*ATLAS*CICT; 920911164227385-CICT-TLSE
Original-To: methods@UK.AC.DARESBURY
Ua-Content-Id: 920911164227385-

Hello netters,
 
Does someone has any experience in direct phage sequencing?
Thank you in advance  for your advices
                      Jacqueline
From: wegloehner@rz-berlin.mpg.de (Wolfgang Wegloehner)
Newsgroups: bionet.molbio.methds-reagnts
Subject: CORRECTION MODIFIED AMINO ACIDS
Date: 4 Sep 92 17:10:44 +0100
Organization: MPI fuer Molekulare Genetik, Berlin

dear net users,

very sorry about a mistake in the previous posting concerning the methylated
amino acids. 

the amino acids we are searching were:

             N(alpha)-trimethyl-L-Alanine
  and        N(epsilon)-trimethyl-L-Lysine


                            Juergen Schmidt
                   MPI fuer Molekulare Genetik, Berlin
From: skrshaw@susssys1.reading.ac.uk
Newsgroups: bionet.molbio.methds-reagnts
Subject: NCTC 1469 cell-line
Date: 12 Sep 92 19:43:06 GMT
Distribution: bionet
Original-To: methods@uk.ac.daresbury

It would seem that bacteria really are intelligent and malicious into 
the bargain. Once again the bugs resident in our incubators have picked 
out the one cell-line for which I have to pay and wiped out both flasks 
of the latest order. Before I go back to the ECACC (British equivalent 
of the ATCC) for yet another lot (65 pounds worth) does anybody have a 
flask they could spare? If you do I would be most grateful!

Sean Shaw
Dept. Microbiology, Reading University
skrshaw@sksscsc1.rdg.ac.uk
Newsgroups: bionet.molbio.methds-reagnts
From: kim@m44.unm.edu
Subject: Salt ppt of RNA
Date: Thu, 10 Sep 92 20:50:05 GMT
Organization: UNM Cancer Center, Albuquerque, NM
Reply-To: kim@m44.unm.edu

Hi group:

I was looking at a BRL/GIBCO catalog and noticed an RNA extraction kit that
claims to yield purified RNA without a phenol extraction step from guanidine
thiocyanate-lysed cells.  A similar kit is offered by the company that makes
the lysate-RNAase Protection Assay (I forget the name).  I was told by the BRL
rep that the procedure takes advantage of selective precipitation of RNA from
GnSCN solutions with high NaCl concentrations.  

Is anyone familiar with this method?  What are the particulars of precipitating
RNA from a cell lysate in GnSCN?  I imagine cells could be lysed in 5M GnSCN,
cell debris spun out, cleared lysate salted, and then the RNA spun out.  The
pellet could then be redissolved and reprecipitated to remove any remaining
DNA, etc., and the RNA either isopropanol precipitated or bound to glass for
final purification and concentration.  This could also be used to purify RNA
probes from their templates after in vitro transcription.

Of course, if hybridization does indeed occur in concentrated GnSCN, as claimed
by Haines and Gillespie (May, 1992 BioTechniques), then it may be possible to
just lyse cells and add oligo(dT) cellulose to get poly(A)+ RNA in one step. 
Has anyone tried that?  I know that it doesn't work in 4M GnSCN, Citrate,
Sarkosyl, Mercaptoethanol (i.e. Solution D), but maybe plain 5M GnSCN/0.1 M
EDTA would be better . . .

Daniel Kim
KIM@FLOVAX.LANL.GOV