HDTenEightyP/NetNews huggingface dataset:
From: ED@micro.uct.ac.za (Ed Rybicki)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: G/C tails in expression constructs
Date: 12 Nov 92 10:14:31 GMT
Reply-To: ed@micro.uct.ac.za
Distribution: bionet
Original-To: rosswhet@edu.ncsu.nrrc.forbt2, methods@uk.ac.daresbury
X-Mailer: Pegasus Mail v2.1c R5.

> To:         methods-and-reagents@net.bio.net
> From:       rosswhet@forbt2.nrrc.ncsu.edu (Ross Whetten)
> Subject:    G/C tails in expression constructs
> Date:       11 Nov 92 18:08:00 GMT

> Hello netters!
> I am making an expression construct with a "constituitive" promoter driving
> transcription of a full-length cDNA that was originally cloned using G/C
> tailing at a Pst I site. Are the G/C tails likely to present problems for
> the expression of the fusion gene? There are likely to be 20 to 50 consecutive
> G residues in the 5' untranslated portion of the resulting mRNA. This is
> a plant construct using the CaMV 35S promoter, if that makes a difference.
> Comments from anyone with experience in this issue are welcome, as are any
> references to papers that address the question. Thanks.

Colleagues of mine had a similar problem with an animal virus gene that
had been cloned using the same method - they simply sequenced it, then
PCRed it out of the clone using primers specific for the 5' and 3' NCR
sequences inside the G/C tails, and recloned it into the expression
vector.  Probably safer, all round.  Hope this is of use.

  ____________________________________________________________________
 | Ed Rybicki, PhD             |    "Now you've got the hang of it    |
 | (ed@micro.uct.ac.za)        | There's nothing you can't do with it |
 | Dept Microbiology           |        If you're very into it        |
 | University of Cape Town     |         You can't go wrong...."      |
 | Private Bag, Rondebosch     |                                      |
 | 7700, South Africa          |               -Mad John              |
 | fax: 27-21-650 4023         | (Ogden's Nut Gone Flake, Small Faces)|
  --------------------------------------------------------------------
From: 76467.702@CompuServe.COM (Alan Gerstein)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Separation Science User Groups
Date: 23 Nov 92 17:23:43 GMT
Distribution: bionet

Hello,
Can you direct me to forums like BIOCSI that put more focus on separation 
techniques like electorphoresis, HPLC et al...?  I've already signed up to the
protein analysis groups within BIOSCI.
Thanks, 
Alan
From: jodi@arbo.microbiol.uwa.oz.au (Jodi Lindsay)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: nonradioactive northerns
Date: 20 Nov 1992 08:00:16 GMT
Organization: The University of Western Australia
Distribution: bionet
NNTP-Posting-Host: cns.microbiol.uwa.edu.au

In article <1992Nov17.142120.26717@gserv1.dl.ac.uk> SHANELT@BIOLAN.UNI-KOELN.DE (Sabine Hanelt) writes:
>From: SHANELT@BIOLAN.UNI-KOELN.DE (Sabine Hanelt)
>Subject: nonradioactive northerns
>Date: 17 Nov 92 14:15:17 GMT



>Hi netters,
>my problem is how to get a nonradioactive northern blot.
>I tried already Digoxigenin DNA and RNA probes (Boeringer System)
>and also biotinylated DNA probes (USB Gene Images). None of these
>systems seem to be helpfull, maybe I make a general mistake.
>Has anyone experiences with nonradioactive northern blots that are not
>so frustrating like mine?

>Thanks for your help
>Sabine



Just a suggestion - have you heard of the Amersham Enhanced 
Chemiluminescence Gene Detection System??  (yes...another kit!!)

It involves labelling probe with horseradish peroxidase using 
glutaraldehyde.  Detection occurs by washing the hybed membrane with luminol 
and peroxidase (and enhancers) which causes a chemical reaction that gives 
off light, which is captured on x-ray film.

Although the standard procedure uses DNA probes it is being successfully 
used with ribosomal RNA in our lab.  The method is fast and sensitive, 
although the detection kits are prone to going off, and are rather 
expensive.  We have not been able to figure out what the "enhancers" are 
so as to make our own.

Regards

Jodi Lindsay
Department of Microbiology
Newsgroups: bionet.molbio.methds-reagnts
From: lhanson@moose.uvm.edu (lhanson)
Subject: Anyone with an IBI Pustell sentinel to sell?
Organization: University of Vermont -- Division of EMBA Computer Facility
Date: Mon, 16 Nov 1992 16:46:55 GMT
From: wetsel_r@wums.wustl.edu
Newsgroups: bionet.molbio.methds-reagnts
Subject: RE: deblocking
Date: 19 NOV 92 16:44:02 GMT
Organization: MIT PLASMA FUSION CENTER
NNTP-Posting-Host: msnews.wustl.edu

In a previous article, obrien@molbio.cbs.umn.edu (Darin O'Brien) wrote:
>I have an oligo given to me by someone else.  Unfortunately, the oligo was not
>deblocked and nobody around here knows how to do it.  Does anyone out there know
>how to do this?  I don't know what solution the oligo is in now.  Can I
>just precipitate the oligo and treat overnight in ammonium hydroxide?  Any help
>would be greatly appreciated.
> 
>darin
> 
-----

Darin:

I assume that the oligo has been deprotected and off the column?  If so, I 
would assume that they are currently in NH4-OH.  Deprotection is then easy. 
We have our oligos in 4ml Wheaton vials available from VWR/Fisher which fit 
nicely into a heating block.  Heat them overnight at 60'C.   It can be done 
in an eppendorf tube as well, in fact there is probably no reason why 
deprotection couldn't be done in a 60'C water bath as long as absolute care 
is taken not to get any water mixed into the oligo.  In the morning, we 
would take 1 ml and then lypholyze it down to dryness, resuspend in 250ul 
water and take OD-260 readings.  Very recently, this net has directed my 
attention to an n-butanol purification/extraction (NAR 19:674) which works 
like a charm!

Good luck and holler if I can help further...

David L. Haviland
haviland@kids.wustl.edu
From: pcovello@ac.dal.ca
Newsgroups: bionet.molbio.methds-reagnts
Subject: Protein phos assays, TCA and dot blot manifolds
Date: 17 Nov 92 18:13:01 GMT
Organization: Dalhousie University, Halifax, Nova Scotia, Canada

Speaking of protein phosphorylation assays, I have a 
question along similar lines. I am looking for a very
rapid method for TCA precipitation/scintillation 
counting of nucleic acids. I have read that a vacuum
manifold can be useful for washing filters and I 
notice that the "Minifold" on p. 20 of the 1992
Schleicher and Schuell catalogue is recommended
for this purpose. Does anyone have any experience
using a vacuum manifold for this purpose. Can anyone
recommend a protocol for its use?

Pat Covello
Dept. Biochemistry
Dalhousie University
Halifax, NS
Canada
From: 76467.702@CompuServe.COM (Alan Gerstein)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Removal of Bind-silane
Date: 20 Nov 92 16:45:46 GMT
Distribution: bionet

Have you tried cycles of scrubbing with lab detergent, followed by rinsing 
with d&d water?
From: 76467.702@CompuServe.COM (Alan Gerstein)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: deblocking
Date: 20 Nov 92 16:45:37 GMT
Distribution: bionet

It might be easier to evaporate the oligo first.  Then, resuspend in 1.0 ml of
concentrated ammonium hydroxide and incubate overnight at 55 degrees C. Use 
hydroxide which had been stored in the freezer, as hydroxide solutions 
evaporate at room temp, and might be too dillute for deblocking.  You can 
perform this in a plastic tube with a screw top cap that contains a gasket, 
but seal with Parafilm too.  You might want to place the tube with the oligo 
inside a scintillation vial containing 2 ml of ammonium hydroxide, cap it and 
seal with Parafilm.  The vapor pressure inside the scintillation vial will 
prevent the tube with the oligo from bursting open. 

Cool down the vials on ice before opening, or the hydroxide will spray all 
over.  You can separate the hydroxide from the oligo via evaporation or gel 
filtration.

Good luck,

Alan
Pharmacia
From: mic268a@vaxc.cc.monash.edu.au
Newsgroups: bionet.molbio.methds-reagnts
Subject: L. lactis plasmid DNA
Date: 20 Nov 92 17:12:10 +1100
Organization: Computer Centre, Monash University, Australia

G'Day netters,

Does anyone out there have a simple procedure for isolating L. lactis plasmid
DNA in reasonable quantities without going through a CsCl gradient?
Are L. lactis plasmids generally high copy or low copy plasmids?

Are there any tricks to treating plasmid DNA from this species, ie
should the DNA be heat treated to inactivate any DNAse etc.?
Any help would be much appreciated.


Thanks in advance


Mike L.
Monash University
Melbourne
AUSTRALIA
From: btozier@pobox.upenn.edu (William A. Tozier)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Need methods for molecular diversity
Date: 19 Nov 92 14:33:24 GMT
Organization: University of Pennsylvania

Hello.
  Recently it has become necessary for us to measure the number of different
compounds in complex mixtures of biological samples. The samples are quite
"messy" from a standard biological point of view, and contain large organic
molecules, RNAs, DNAs, and proteins. We've embarked on a multistep process
that involves chromatography, electrophoresis, and so on, but since these
techniques are mainly optimized for purification and not "peak-counting",
we're skeptical that we'll be able to discriminate "how many kinds of
things" are in a given sample. We know there are over a thousand, maybe
millions. Any supportable estimate is all we're looking for.
  Since this will end up being a survey of several mixtures, we'd really
appreciate some help. Are we missing some elementary technique that might
simplify our task? Something perhaps obsolete for most people that might
work here?
  As a specific question: Is anyone aware of an electrophoretic technique that
would discern DNA, RNA and protein simultaneously?

Thanks for your help,
Bill Tozier
Biology Department
University of Pennsylvania

btozier@pobox.upenn.edu
bill@amy.med.upenn.edu
From: gamel@rzmain.rz.uni-ulm.de (Anton J. Gamel)
Newsgroups: bionet.molbio.methds-reagnts
Subject: LINSCOTTS DIRECTORY -- real address
Date: 22 Nov 92 15:44:07 GMT
Distribution: bionet
Original-To: "methods@daresbury.ac.uk"@1.decnet

Hello folks!

Some weeks ago I posted the corrected address (after move) of
LINSCOTTS DIRECTORY of immunological & biological Reagents,
CALIFORNIA.  Now it is clear why I had to wait for an answer:
This oversea settlement redirected the letter to the firm in GB.
Here the address:

LINSCOTTS DIRECTORY
PO BOX 55
EAST GRINSTEAD
SUSSEX RH19 3YL
GREAT BRITAIN

TEL: 0342 82 4854
FAX: 0342 82 4212

The reference is: J.Imm.Meth. 120, 145 (1989)

    Anton J. Gamel

If possible please reply directly to <GAMEL@DULRUU51.bitnet>
+-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-+
:                                : <GAMEL@DULRUU51.bitnet>           o o    :
:         Anton J. Gamel         : <GAMEL@RZMAIN.RZ.UNI-ULM.DE>       V     :
:     -  -  -  -  -  -  -  -     : <GAMEL@PATVCL.RZ.UNI-ULM.DE>     (,,)    :
:       Look, a Vet on VAX       : <GAMEL@RZ.UNI-ULM.DBP.DE>        -""--   :
:                                : <GAMEL@INFORMATIK.UNI-FREIBURG.DE>       :
+-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-+
From: wetsel_r@wums.wustl.edu
Newsgroups: bionet.molbio.methds-reagnts
Subject: Immunoprecipitation question...
Date: 23 NOV 92 15:59:40 GMT
Organization: MIT PLASMA FUSION CENTER
NNTP-Posting-Host: msnews.wustl.edu

Greetings Netters:

We do immunoprecipitations all the time in our lab, both biosynthetically
labeled and surface labeled, with polyclonal and monoclonal antiobodies,
respectively. 

My question is:  Does anyone have a protocol for doing immunoprecipitations
from biosynthetically labeled cells with monoclonals? 

Our polyclonal protocol works great but uses 0.5 Triton X100, 0.5% DOC, and
1% SDS but as one might expect this wash solution doesn't cut it for
monoclonals, in fact, the SDS blows them away leaving me with a blank gel! 
Decreasing the SDS, so far, only increases the background.  Has anyone
tried various salt combinations? 

Any suggestions?

Thanks in advance,

David L. Haviland
haviland@kids.wustl.edu
Newsgroups: bionet.molbio.methds-reagnts
From: erman@binah.cc.brandeis.edu
Subject: in vivo crosslinking
Reply-To: erman@binah.cc.brandeis.edu
Organization: Brandeis University
Date: Sat, 21 Nov 1992 18:18:45 GMT

Does anyone know of any references (or methods) that enable one to 
put BUDR or any other crosslinking agent into mammalian cells in order
to crosslink protein to DNA in vivo. Any ideas about the efficiency of 
crosslinking and whether I'd be able to fish out specific protein-DNA
complexes, say with a specific probe to the binding site and an antibody
to the suspected protein?
Batu ERman
From: pedykema@iastate.edu (Philip E Dykema)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Help needed: genomic DNA isolation
Date: 18 Nov 92 00:27:55 GMT
Organization: Iowa State University, Ames, IA

I can relate to having problems isolating good quantities of plant genomic DNA

Here is the method I presently use:
1:CTAB Buffer = 2%CTAB/100mMTris pH 8/20mMEDTA/1.4MNaCl/0.2%BMEtoh/1%PVP-40;
   grind 2-5g leaves under liquid nitrogen; add to buffer in a 1:3 ratio;
   incubate for 1 hour.
2:0.6x ice-cold isopropanol; pellet in clinical centrifuge.
3:Resuspend in 0.5-1.0 ml TE
4:Extractions = phenol/chloroform 1x/ether 1x/chloroform 1x.
5:Ethanol precipitate
6:Resuspend in 1.0 ml TE (I use 65C to help both resuspensions; it helps 
   unwind the DNA strands and also eliminates DNAse contaminations)
   At this point I have ~200 - 800 ug DNA
7:Proceed with a CsCL centrifugation.

________________
Philip Dykema (pedykema@vincent.iastate.edu)
Botany Department
Iowa State University
From: ca566@cleveland.Freenet.Edu (Michael Holloway)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: seq. gel problems
Date: 21 Nov 1992 18:05:43 GMT
Organization: Case Western Reserve University, Cleveland, OH (USA)
Reply-To: ca566@cleveland.Freenet.Edu (Michael Holloway)
NNTP-Posting-Host: hela.ins.cwru.edu


In a previous article, adeno@dbe.csiro.au (both lab) says:

>We are having problems with sequencing gels lately, we see a lot of smearing
>or patches where the ladder is fuzzy.  Any suuggestions ?
>Thanks for your advice.
>Adam
>

High background kind of smearing or are the banks just distorted?  If the
latter then I would predict that it has something to do with pouring the
gel.  If the former then it has got to be a template contamination problem
(assuming that the kit is still good).
-- 
E-mail: mhollowa@ccmail.sunysb.edu (mail to freenet is forwarded)
phone:  (516)444-3612
Newsgroups: bionet.molbio.methds-reagnts
From: bhjelle@carina.unm.edu ()
Subject: Re: PCR Mutagenesis
Date: Mon, 16 Nov 92 14:43:46 GMT
Organization: University of New Mexico, Albuquerque

In article <Bxntyw.Fqu@usenet.ucs.indiana.edu> jgraham@bronze.ucs.indiana.edu (the End) writes:
>In <mxdq3j#@lynx.unm.edu> bhjelle@carina.unm.edu () writes:
>
>>I encountered an impressively high level of mutagenesis (14 
>>errors out of 411bp) in amplifying a segment of DNA from
>>a plasmid clone. All mutations involved T or A in some
>>manner. 11 were point mutations (transitions and
>>transversions), but 3 were 1 bp insertions or deletions.
>
>Very interesting and disturbing. We routinely generate templates for in vitro
>transcription via PCR using a protocol (Higuchi et al. NAR 16 (15) p.7351) 
>recomended by a well-known colleague.
>
>This procedure uses plasmid targets at 200-400 ng/ rxn. with dNTPs at 0.175 mM
>and a mere 16 cycles. The low cycle number is suggested in order to 
>reduce the chance of errors generated by Taq. I obtain about 1 to 5 ug of
>500 bp fragments from target plasmids around 3 Kb.
>
>In the NAR paper describing this technique, Krummel and associates report that
>they have not seen a single base change in preparing 10 different transcription
>templates in the 500 bp size range (p. 7359 line 14).
>
>Are you using a significantly lower dNTP concentration ?
>
I used 0.8mM of (each) dNTP, 4mM MgCl2 (these primers were optimized
at those concentrations) and 250 pmol of primer (each) in TV=125ul.
I got a high yield (5ug) of PCR product, but in back-calculating,
it seems that I would have gotten 250ug of product if all
dNTPs in solution were really "used up" in the reaction. So
I have a hard time believing that Taq ran out of dNTPs during
PCR. However, I did use 45 cycles for convenience.

At the suggestion of the P-E rep, I lowered the dNTPs to 200uM,
the Mg to 1.5mM, and did 10, 15, and 20 cycles to regenerate
PCR product. These parameters should all be lower to generate
less mutations. Also, I previously used 12.5U of AmpliTaq but
now lowered that to 2.5U. I used 1ug of plasmid template(!).

Hopefully I will be able to get back with results showing lower
levels of mutations.

Brian
From: DMEIER@mis.mcw.edu
Newsgroups: bionet.molbio.methds-reagnts
Subject: Peptide affinity columns
Date: 16 Nov 92 22:33:52 GMT
Distribution: bionet
Organization: Medical College of Wisconsin (Milwaukee, WI)

Dear Netters,
  I plan to have a peptide (10-mer) with a cysteine on the amino 
terminal end so I can link it to BSA for injection into rabbits.  Is 
there an amino acid residue that should not be next to the cysteine, 
or does it matter.  I also plan to set up a peptide affinity column 
later to affinity purify my antibody.
  Any advice would be gratefully received.

Dan Meier
Medical College of Wisconsin
DMEIER@mis.mcw.edu
From: BEMDAM@rulgl.LeidenUniv.nl (EDWIN TEN DAM)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Opinions on TnT Lysate ?
Date: 18 Nov 92 16:08:00 GMT
Distribution: bionet
X-Envelope-To: methods@daresbury.ac.uk
X-Vms-Cc: BEMDAM
Original-To: methods@uk.ac.daresbury
X-Vms-To: IN%"methods@daresbury.ac.uk"

Hi Netlanders!

Has anyone outhere used the TnT Lysate System from Promega? Yes, it's a kit,
8-) but I guess most of you don't make rabbit reticulocyte lysates yourselves.
TnT is a system for coupled transcription/translation. It's available in T7 and
SP6 flavours, and it's big advantage <at least on paper> is that you can use
miniprep plasmid DNA as a template. No digestions, no cleaning up your RNA etc.
Sounds pretty good. And fast.
I'd like to know your experiences, especially with respect to efficiencies and
pricing versus separate transcription/translation.

Thanks, Edwin

If i get a lot of responses, I'll summarise. No, I don't work for Promega._

Edwin ten Dam			Internet:  bemdam@rulgl.leidenuniv.nl
Dept. of Biochemistry		phone:	   (31)71-274294
Gorlaeus Labs			FAX:	   (31)71-274340
Leiden University
PO Box 9502
2300 RA Leiden
The Netherlands
From: ca566@cleveland.Freenet.Edu (Michael Holloway)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Beta-gal cotransfection - nonsense results?
Date: 23 Nov 1992 18:29:09 GMT
Organization: Case Western Reserve University, Cleveland, OH (USA)
Reply-To: ca566@cleveland.Freenet.Edu (Michael Holloway)
NNTP-Posting-Host: hela.ins.cwru.edu



Does it make any sense to anyone that Beta-gal assay results from
cotransfected cell extracts should vary with the size of the test CAT
plasmid?  Several weeks ago I obtained that result.  As the size of my
upstream region decreased in a nest set of CAT reporters, the Bgal numbers
went down till there was a factor of 10 difference between the highest and
lowest.  My best guess was that there was too much plasmid in the
electroporation and that I would have to optimize.  

Now I see in this last issue of Biotechniques (13:700) a table of CAT and
Bgal results that look exactly like mine.  As the CAT plasmid changes so
does the Bgal result.  When the CAT value is normalized the progression
shown by the CAT results is turned completely around (the highest CAT value
becomes the lowest CAT value).  This makes no sense to me.  Can someone
offer any explaination for why there should be a progressive change in Bgal
values?  If this takes place commonly how can one justify the use of Bgal
to normalize reporter values?

Puzzled
Mike
-- 
E-mail: mhollowa@ccmail.sunysb.edu (mail to freenet is forwarded)
phone:  (516)444-3612
From: fyamao@ddbj.nig.ac.jp (Fumiaki Yamao)
Newsgroups: bionet.molbio.methds-reagnts
Subject: where is vecbase?
Date: 17 Nov 92 04:20:58 GMT
Organization: National Institute of Genetics in Japan

Recently I learned database of vector sequences, VECBASE. How can I access
to this database? Or, Can I get this file via FTP. Any infomation or 
suggestions would be appreciated.

Thanks in advance.

--
Fumiaki Yamao                     fyamao@ddbj.nig.ac.jp
National Institute of Genetics    Phone:+81-559-72-2732
Mishima 411, Japan                  Fax:+81-559-75-6240
From: ccy@po.CWRU.Edu (Cheung C. Yue)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Wanted: practical experience with Gene Sprinter
Date: 15 Nov 92 15:50:29 GMT
Reply-To: ccy@po.CWRU.Edu (Cheung C. Yue)
Organization: Case Western Reserve University, Cleveland, OH (USA)
NNTP-Posting-Host: slc5.ins.cwru.edu


I saw the video from Fotodyne on their new sequencing package called 
Gene Sprinter.  It basically uses 50 micron thick acylamide gels run
in a controlled-temperature apparatus using a 10,000 volt power supply.
Gel runs are typically 30 minutes or so.
Gels are dried right on the glass plate by a simple heating device with
no need for vacuum.  Volumes of running buffers used amount to less than
100 or so ml. per run.  The gels are a bit short so they claim readings
of 250 bp or so.  From my perspective, this apparatus seems ideal for
many of the shorter sequencing runs one needs in a mol biol lab.  The
price tag for the whole package amounts to $10,000 or so.  They told
me they would not sell any components of the systems separately (kind
of wasteful in my case since I have 2 recirculating pumps in the lab 
already).  I am curious if anyone on the net have had direct practical
experience with this system, and whether things are as easy and convenient
as they seem in the demo.

I have no connections with Fotodyne.  I am merely interested in feedback
on this system.

ccy@po.cwru.edu
-- 
Xref: sparky bionet.molbio.methds-reagnts:2606 bionet.molbio.gdb:23 bionet.molbio.genbank:535
From: shahn@hstbme.mit.edu (Samuel Hahn)
Newsgroups: bionet.molbio.methds-reagnts,bionet.molbio.gdb,bionet.molbio.genbank
Subject: Looking for sequences for ...
Date: 23 Nov 92 04:38:22 GMT
Reply-To: shahn@hstbme.mit.edu (Samuel Hahn)
Organization: Massachusetts Institute of Technology

I was wondering if anyone out there can help me. I am looking for the cDNA
sequences for some human genes, specifically the LDL receptor gene, the
apolipoprotein B-100 gene and the apolipoprotein E gene (E3 allele).

If anyone happens to have these sequences around in a convenient emailable
format, I would certainly greatly appreciate it! Alternatively, if someone can
tell me how to get them myself, I would appreciate that just as much. I thought
that there was an anonymous FTP site for DNA sequences at NIH but I can't seem
to navigate my way through the directories to get the sequences that I am
interested in.

Please email replies to "shahn@hstbme.mit.edu" as I do not regularly read 
these newsgroups. Thanks in advance.

Sam
Newsgroups: bionet.molbio.methds-reagnts