HDTenEightyP/NetNews huggingface dataset:
From: chai_z@wehi.edu.au
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: lambda won't cut
Date: 14 Sep 92 01:18:36 GMT
Distribution: bionet
Organization: Walter & Eliza Hall Institute

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


I had the same problem before. What I am doing now is to use DNA grade agrose
to make media plate and the soft top agrose to grow my lambda phages, rather
than the normal agar. The cell lysates are collected in SM buffer, discard the
cell debris by low speed centrifugation. The virus was pelleted by
centrifugation with PEG. Then, resuspended in TE, phenol-chlroform and
chloroform extraction. Finally, ethanol precipitated and resuspended in TE.
This worked well for me for all the lambda clones I have handled.

Important thing is the normal bacterial media agar may contain something 
inhibiting the restriction enzymes. refer to the Maniatis' Molecular Cloning.
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Re: ASAP
From: cytogen@chmeds.ac.nz (Martin Kennedy)
Date: 11 Sep 92 14:29:21 +1200
Distribution: bionet

In article <009605D8.788A4200.383@NTBTOX.NCTR.FDA.GOV>, wmelchior@NTBTOX.NCTR.FDA.GOV ("Bill Melchior, NCTR/FDA") writes:
>>Yes, there really is an Artic shrimp alk. phos. [ASAP]
>>....Even more suprising is that
>>it really works.  Best part is that you can digest and SAP at the
>                                              ^^^^^^^^^^^^^^
>>same time.
> 
> Do you mean restriction digest and dephosphorylate simultaneously?
> While typical restrictions are done near neutrality (pH 7.5 for the Maniatis
> etal buffers), USB compares ASAP with calf intestinal AP at pH 9.6.
> (But they do suggest pH 8 for the reaction -- why?)  Have you used ASAP with 
> different restriction buffers?  Could you be specific about your conditions?
> 
> Thanks.
> ________________________________________________________________________________
> The opinions stated are mine, not those of NCTR or its sponsoring organizations.
> 
> Bill Melchior                                ||   "You have lawyers the way
> National Center for Toxicological Research   ||    other people have mice."
> Jefferson, AR  72079                         ||
> (501) 543-7206                               ||   -Kenneth Duncan, English
>                                              ||    Health & Safety Executive,
> WMELCHIOR@NTDOC.NCTR.FDA.GOV                 ||    to US regulators


I use conventional Calf Alkaline Phosphatase (from Boehringer), and always
add it directly into my restriction digestions about an hour before completion
of the digestion and without changing the buffer.  Boehringer put out a
protocol describing this method years ago (ca 1986).  I've used it since then
without any major problems.  What's more, you can heat inactivate this, albeit
                                              ^^^^^^^^^^^^^^^^^^^
at 75o for 15 minutes, and I don't bother phenol extracting or precipitating
vectors prepped in this way - it works, at least most times, believe it or not.
From: sem@cxa.daresbury.ac.uk ("S. Marshall")
Newsgroups: bionet.molbio.methds-reagnts
Subject: ftp FAQ files
Date: 8 Sep 92 09:18:31 GMT
Reply-To: sem@cxa.daresbury.ac.uk
Distribution: bionet
Original-To: methods@uk.ac.daresbury


	From rmkdhhp@ucl Mon Sep  7 18:39:07 1992
	Received: from dlgmp.dl.ac.uk by uk.ac.dl.cxa; Mon, 7 Sep 92 18:39:05 +0100
	Reply-To: rmkdhhp@ucl
	Via:   UK.AC.BCC.MAIL-A;  Mon, 7 Sep 92 18:37 WET (V41 at UK.AC.DARESBURY.DLGM)
	Received: from link-1.ts.bcc.ac.uk by mail-a.bcc.ac.uk with SMTP (PP)
	          id <12336-0@mail-a.bcc.ac.uk>; Mon, 7 Sep 1992 18:35:52 +0100
	From: Dr Huaizheng Peng <rmkdhhp@ucl>
	Message-Id: <110451.9209071735@uk.ac.bcc.ts>
	To: bionet-news@daresbury
	Date: Mon, 07 Sep 92 18:35:43 +0100
	
	Can you tell me where I can ftp FAQ files about molecular biology methods?
	
	Thank you very much.
	
	Dr Peng HZ
	Dept. of Histopathology
	UCH, London
	
Newsgroups: bionet.molbio.methds-reagnts
From: jgraham@sunflower.bio.indiana.edu (Jim Graham)
Subject: ssDNA sequencing preps: NaOAc or NOT ?
Followup-To: bionet.molbio.methds-reagnts
Organization: Indiana University
Date: Tue, 8 Sep 1992 23:53:18 GMT

Maybe it's a worn thread but :

I recently have been using what was describe in a bulletin from Stratagene
as an improvement in the organic extraction procedure for cleaning up
phagemid templates. I am using it with Sequenase single stranded
sequencing. They claim that a SINGLE phenol-CIA extraction is sufficient if
the PEG phage pellet from a 1.5 ml culture supernatant is resuspended in a
400ul of  0.3 M NaOAc. Others in my lab agree, but I find that more than
half of my samples show a large grey smear instead of a ladder. Must I
return to the laborious 2 phenol/CIA, 3 CIA extraction procedure ? I have
always found that to work, but it is a real pain in the @#$#.

Thanks for your help,

Jim
J. E. Graham
Biology/Chemistry Departments
Indiana University -Bloomington
.............."Il n'y a pas des sciences apliquees, mais il y
a..............
.................des applications de la science". L.
Pasteur.................
From: skspoidn@susssys1.reading.ac.uk
Newsgroups: bionet.molbio.methds-reagnts
Subject: HgiE II
Date: 8 Sep 92 10:02:14 GMT
Distribution: bionet
Original-To: methods@uk.ac.daresbury

Thanks to the (now) 3 people that responded to my Q.

Mike
From: SMITH_DENNIS_P@LILLY.COM ("DENNIS P. SMITH")
Newsgroups: bionet.molbio.methds-reagnts
Subject: UV Shadowing
Date: 10 Sep 92 20:22:15 GMT
Distribution: bionet

Need advice of how to take photograph of an oligonucleotides using UV
shadowing. Can anyone suggest a reliable technique? Thanks DPSMITH


From: SMITH DENNIS P                (MCVAX0::BOOKS)

To:   VMS MAIL ADDRESSEE            (INT::"methods@genbank.bio.net")
From: BECKERM@SIMSC.EARN
Newsgroups: bionet.molbio.methds-reagnts
Subject: AMMONIA CLEAVAGE FROM PROTEINS
Date: 14 Sep 92 16:33:00 GMT
Distribution: bionet
X-Original-To: METHODS@DARESBURY.AC.UK, BECKERM
Original-To: METHODS@UK.AC.DARESBURY

I'M LOOKING FOR REFERENCES OF AMMONIA CLEAVAGE FROM PROTEINS, PARTICULARLY
LIGHT INDUCED MECHANISMS AND ARE SOME AMINO ACIDS MORE SUSCEPTIBLE THAN
OTHERS.  ALSO INTERESTED IN FREE RADICAL FORMATION IN THE SOLID STATE AND
POSSIBLE FORMATION OF ARTIFACTS UNDER NORMAL ACID HYDROLYSIS CONDITIONS
(INCLUDING AMMONIA RESULTING FROM THE FREE RADICALS)
        I'D APPRECIATE ANY INFORMATION, MARY
(BECKERM@MSCSI.BITNET)
From: rbonfigl@waite.adelaide.edu.au (Rod Bonfiglioli)
Newsgroups: bionet.molbio.methds-reagnts
Subject: expression vectors for proteins
Date: 13 Sep 92 21:32:25 GMT
Distribution: bionet

does anyone have any experience of using expression vectors for largish
proteins, i am trying tofind one that will take inserts of up to 3 Kb to
express proteins of the 100 Kd range.  i have tried pGEX vectors, and they
dont seem to want do, anyone got any suggestions please?

thanx in advance

rod bonfiglioli
email rbonfigl@schooner.waite.adelaide.edu.au
From: c50951@sakura.kudpc.kyoto-u.ac.jp (yukio nagano)
Newsgroups: bionet.molbio.methds-reagnts
Subject: lovastatin
Date: 7 Sep 92 03:21:19 GMT
Organization: Department of Food Science and Technology, Kyoto University, Kyoto, Japan
Originator: c50951@sakura.kudpc.kyoto-u.ac.jp

Dear bio-netters,

Please tell me how to get a lovastatin, an inhibitor of hydroxymethylglutaryl-
coenzyme A reductase.

--
/* Yukio Nagano, Department of Food Science and Technology,                */
/*               Faculty of Agriculture, Kyoto University                  */
/* email: c50951@sakura.kudpc.kyoto-u.ac.jp                                */
Organization: University of Maine System
Date: Wednesday, 9 Sep 1992 11:05:21 EDT
From: Robert Roxby <RROXBY@MAINE.MAINE.EDU>
Newsgroups: bionet.molbio.methds-reagnts
Subject: Quantitative Autoradiography???

I saw a discussion somewhere recently about the pitfalls of
using autorads of blots, etc., to estimate quantities of
things, eg message levels.  I dont think it was a regular
publication, but it had references.  I have lost this reference
and will appreciate it if anyone can put me bace onto it.
Thank you.
From: sbhattac@crc.ac.uk (Dr. Shoumo Bhattacharya B1 061)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Epstein Barr
Date: 8 Sep 92 21:38:21 GMT
Distribution: bionet
Original-To: methods@uk.ac.daresbury

Just a comment.

Epstein Barr virus is used to 'immortalize' lymphoblastoid cells from 
humans. Avoid transforming cell lines of laboratory staff as accidental
self inoculation with your own immortalised cell line will not be a 
good thing.

Dr. Shoumo Bhattacharya				
MRC Molecular Medicine Group
Royal Postgraduate Medical School
Hammersmith Hospital, DuCane Road London W12 0NN
Newsgroups: bionet.molbio.methds-reagnts
From: Klaus.Matthaei@anu.edu.au
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: ssDNA sequencing preps: NaOAc or NOT ?
Date: 9 Sep 92 02:14:47 GMT
Distribution: bionet

>Maybe it's a worn thread but :
>
>I recently have been using what was describe in a bulletin from Stratagene
>as an improvement in the organic extraction procedure for cleaning up
>phagemid templates. I am using it with Sequenase single stranded
>sequencing. They claim that a SINGLE phenol-CIA extraction is sufficient if
>the PEG phage pellet from a 1.5 ml culture supernatant is resuspended in a
>400ul of  0.3 M NaOAc. Others in my lab agree, but I find that more than
>half of my samples show a large grey smear instead of a ladder. Must I
>return to the laborious 2 phenol/CIA, 3 CIA extraction procedure ? I have
>always found that to work, but it is a real pain in the @#$#.
>
>Thanks for your help,
>
>Jim
>J. E. Graham
>Biology/Chemistry Departments
>Indiana University -Bloomington
>.............."Il n'y a pas des sciences apliquees, mais il y
>a..............
>.................des applications de la science". L.
>Pasteur.................


I have always used one phenol extraction then one IAC extraction, never
twice with excellent results.  BUT the major problem is with the removal of
the PEG.  ALWAYS aspirate, then briefly respin and re-aspirate to dryness. 
The PEG causes major problems with sequencing.

Hope that this helps.

Cheers, Klaus. 
--------------------------------------------------------------------------
Klaus Matthaei
Gene Targeting
The John Curtin School of Medical Research
The Australian National University
E-mail: Klaus.Matthaei@anu.edu.au

"Don't try and fix it if it ain't broke".
--------------------------------------------------------------------------
From: wetsel_r@wums.wustl.edu
Newsgroups: bionet.molbio.methds-reagnts
Subject: Ligation trouble...
Date: 8 SEP 92 12:31:38 GMT
Organization: MIT PLASMA FUSION CENTER
NNTP-Posting-Host: msnews.wustl.edu

Hello Netters:

I have a old problem.  That being trouble with a ligation.  Most of the time
we don't encounter too much trouble; however, not this time.  The only enzyme
we could use for this ligation was SphI which leaves 3' overhangs.  The
ligations have been giving us fits.  Vectors, pSP72 & pGEM4, have been cut,
alkaline phosphatased, and even put through mock ligations (no insert) and all
run at around the correct length for the cut vector.  We are currently 0-8 in
the ligation game so far.  We don't have that much experience with this
enzyme... is there something simple that we're leaving out in trying to do
this ligation???????

Thanks in advance.

David L. HavilandADD/EDIT
haviland@kids.wustl.edu
(stumped in SphI ligations)

Newsgroups: bionet.molbio.methds-reagnts
From: broe@aardvark.ucs.uoknor.edu (Bruce Roe)
Subject: Re: UV Shadowing
Originator: usenet@kittyhawk.ecn.uoknor.edu
Date: Sun, 13 Sep 1992 11:58:00 GMT
News-Software: VAX/VMS VNEWS 1.41    
Organization: University of Oklahoma - University Computing Services

In article <18qm5pINNscj@usenet.INS.CWRU.Edu>, djt2@po.CWRU.Edu (Dennis J. Templeton) writes...
> 
>In a previous article, SMITH_DENNIS_P@LILLY.COM ("DENNIS P. SMITH") says:
> 
>>Need advice of how to take photograph of an oligonucleotides using UV
>>shadowing. Can anyone suggest a reliable technique? Thanks DPSMITH
> 
>Put your gel on a piece of Saran Wrap, and lay it atop a fluorescing
>substrate (More on this below).  Then use a hand illuminator to shine
>through the gel, and detect dark spots on the otherwise bright background.
> 
>Finding the right fluorescing plate is sometimes tricky.  What always works
>is the glass cellulose TLC plates that are marked "with fluorescence
>indicator"; maybe the plastic plates or the silica TLC plates work too.  We
>use instead an intensifying screen, but it turns out that many screens
>don't work (they don't fluoresce brightly)... we tried about 15 and only
>one (an old one with lots of cracks) works for us.  
> 
>If you use the glass fluorescing plates, you can put the gel on a
>transilluminator, cover with Saran, then place the plate over and view from
>the top, i.e. the back of the plate.
> 
>good luck,  dennis

We use a similar procedure with plastic-backed silica TLC plates "with
fluorescence indicator and they work fine.
-- bruce
  - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
 \  Bruce A. Roe                     Dept. Chemistry and Biochemistry /
 /  BROE@aardvark.ucs.uoknor.edu     University of Oklahoma           \
  - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
Wed, 9 Sep 1992 12: 15:21 +0200
               Wed, 9 Sep 1992 15: 21:14 +0200
From: suter@VAX.MPIZ-KOELN.MPG.dbp.de
Newsgroups: bionet.molbio.methds-reagnts
Subject: sph1
Date: 9 Sep 92 13:21:14 GMT
Distribution: bionet
Content-Identifier: 12
Original-To: methods@uk.ac.daresbury
Conversion: Prohibited




sphI is a enzyme which leaves 3' overhangs and is very hard to cip-treat.

clemens
From: LLR8156%USA.decnet@USAV01.GLAXO.COM ("USA::LLR8156")
Newsgroups: bionet.molbio.methds-reagnts
Subject: re: cDNA library kits
Date: 4 Sep 92 22:40:00 GMT
Distribution: bionet


     I have had InVitrogen make a cDNA library for me recently, and was
very pleased with the result.  The library they constructed had four million
members.  The cost is $2000 if you send them the mRNA, for an additional
$1000, they will isolate the mRNA for you.  Another colleague had two
libraries made recently (three million members from one, seven from the other).
InVitrogen sells the same kits they use to make their libraries, however,
if you only have four months to construct and screen it, it might be advantegous
to have it made.  Several biotech companies are selling premade libraries from
different tissues, and ATCC has some libraries as well-it might be good to
see if what you need is already available.  Another possibility might be to
check the literature and see if someone has made a library from the same 
tissue-typically you amplify the library and freeze down aliquots. Maybe
they would be willing to send you an aliquot.
     Best wishes;
     Lisa Ross
     Glaxo Research Institute, Inc.
P.S. No connection to In Vitrogen - I just buy their products.
From: bsh@MED.PITT.EDU (Basavaraju Shankarappa)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Tacking down cells
Date: 8 Sep 92 23:51:08 GMT
Distribution: bionet

> >Hello netters,
> >I recently read somewhere about a product that will "tack down"
> >non adherant cell to a dish or plate. Does anyone know of this product. Where 
> A product named "cell tack"(?) used to be (4-5 years ago) advertised in the
> back pages of the popular journals.  It was essentially barnacle glue, and
> was most useful to leukocyte biologists, I think.  I can't recall the
> company name, and I havent seen it advertised recently.  Any more help out
> there?

As I understand, special (!) flasks that have been coated with antibodies
specific to certain classes of leukocytes like that for CD8 have been/are 
being used for isolation of the CD8 cells and other similar applications.

I know of people who are using this, but I donot know the company name
and other details.  If anybody is interested in knowing these details
please post and I will provide the details.

Raj Shankarappa
Univ of Pittsburgh
From: MBWEN@CCVAX.AS.EDU.TW
Newsgroups: bionet.molbio.methds-reagnts
Subject: commercial cDNA library
Date: 16 Sep 92 01:17:00 GMT
Distribution: bionet

I am interested in purchasing a Chinese hamster ovary (CHO) cDNA library. Does anyone know which company makes better cDNA library? Strategene, Clontech, or others? Thanks in advance.

Wen Chang
IMB, Academia Sinica, 
Taipei, Taiwan
E-mail address: IN%"mbwen@ccvax.as.edu.tw"
Fri, 11 Sep 1992 12: 06:33 +0100
               Thu, 10 Sep 1992 15: 43:50 +0100
               Thu, 10 Sep 1992 18: 49:39 +0100
From: suter@VAX.MPIZ-KOELN.MPG.dbp.de
Newsgroups: bionet.molbio.methds-reagnts
Subject: re: some more contaminants
Date: 10 Sep 92 17:49:39 GMT
Distribution: bionet
Content-Identifier: 12
Original-To: methods@uk.ac.daresbury
Conversion: Prohibited

Mike Grosse wrote:

<Subj:	lambda won't cut
<
<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

i had EXACTLY the same problem two months ago: same enzyme, same technique,
same results ! my boss suggested that there is something in the LB/top agarose
that inhibits digestion, and you won't be able to get rid of it with 
standard procedures (heavy metal ? polysaccharides ? i don't know). anyway,
these same compounds may trouble you when you ligate in low melting agarose
also...

after many different purifications, all in vain, i purified the DNA
over a QIAGEN column, and presto: perfect DNA, perfect Eco digest, perfect
ligation.

if you don't have these kind of columns, perhaps you should use another
kind that is used for DNA preparation in your lab, i just followed
the dna miniprep protocol (perhaps sephadex would also work, altho the principal
is completely different).

good luck !

clemens
----------------------------------> cut here <----------------------------------
Clemens Suter-Crazzolara, PhD
Max-Planck-Institut fuer Zuechtungsforschung
Abteilung Genetische Grundlagen der Zuechtungsforschung
Carl-von-Linne Weg 10
5000 Koeln 30
Tel. xx49-221-5062.221           fax. xx49-221-5062.21
suter@vax.mpiz-koeln.mpg.dbp.de
------------------------------------------------------------------------------- 
     ! all job offers should include a self-addressed, stamped envelope !
-------------------------------------------------------------------------------
From: robert@arbo.microbiol.uwa.oz.au (Robert Coelen)
Newsgroups: bionet.molbio.methds-reagnts
Subject: re: 2809   re: Epstein Barr   (Anthony Davies)
Date: 11 Sep 1992 09:15:43 GMT
Organization: University of Western Australia
NNTP-Posting-Host: mve.microbiol.uwa.edu.au


>                                       |||One of the Klein's worked with EBV for a
>> lifetime and never seroconverted !|||
>
>Actually Eva Klein did convert a few years ago - - from her granddaughter!!
From: rbonfigl@waite.adelaide.edu.au (Rod Bonfiglioli)
Newsgroups: bionet.molbio.methds-reagnts
Subject: correction to PCR message
Summary: correction to my last pcr message
Keywords: correction to pcr
Date: 15 Sep 92 06:32:49 GMT
Organization: The University of Adelaide, Waite Campus
NNTP-Posting-Host: schooner.waite.adelaide.edu.au
Faculty: Agricultural & Natural Resource Sciences

i think i said in the PCR message just before this, that i was trying to join
together fragments of 100-200nts, this should read 1000-2000nts ta, rod
From: bsh@MED.PITT.EDU (Basavaraju Shankarappa)
Newsgroups: bionet.molbio.methds-reagnts
Subject: Re: Re: ASAP
Date: 10 Sep 92 14:58:34 GMT
Distribution: bionet

> >Yes, there really is an Artic shrimp alk. phos. [ASAP]
> >....Even more suprising is that
> >it really works.  Best part is that you can digest and SAP at the
> >same time.
> Do you mean restriction digest and dephosphorylate simultaneously?
> While typical restrictions are done near neutrality (pH 7.5 for the Maniatis
> etal buffers), USB compares ASAP with calf intestinal AP at pH 9.6.
> (But they do suggest pH 8 for the reaction -- why?)  Have you used ASAP with 
> different restriction buffers?  Could you be specific about your conditions?
> 
> Thanks.
> 
> Bill Melchior                                ||   "You have lawyers the way
> National Center for Toxicological Research   ||    other people have mice."
> Jefferson, AR  72079                         ||
> (501) 543-7206                               ||   -Kenneth Duncan, English
>                                              ||    Health & Safety Executive,
> WMELCHIOR@NTDOC.NCTR.FDA.GOV                 ||    to US regulators
> 

The technical folk at New England Biolabs basically suggested that you can
use their CIAP with any of their restriction enzyme buffers.  I have been 
doing this with considerable if not absolute success.
Raj Shankarappa
Univ of Pittsburgh.

Newsgroups: bionet.molbio.methds-reagnts
From: jgraham@bronze.ucs.indiana.edu (the End)
Subject: Re: ssDNA sequencing preps: NaOAc or NOT ?
Organization: Indiana University
Distribution: bionet
Date: Wed, 9 Sep 1992 20:24:14 GMT


Although many feel that residual PEG is a big deal in preparing single
stranded sequencing templates, Pheiffer and Zimmerman (NAR 11(2)p.7853
1983) report that 95% of the PEG 6000 from a 15% solution is removed
from the aqueous phase by a single extraction with 3 vol. chloroform
at room temp. It seems then quite unlikely that PEG would contaminate
sequencing templates prepared with traditional organic extractions.
Perhaps this belongs in Ed's urban myth collection. Thanks for the
tips regardless.


Jim
Newsgroups: bionet.molbio.methds-reagnts
From: broe@aardvark.ucs.uoknor.edu (Bruce Roe)
Subject: Re: TYE Medium?
Originator: usenet@kittyhawk.ecn.uoknor.edu
Date: Sun, 13 Sep 1992 12:09:00 GMT
News-Software: VAX/VMS VNEWS 1.41    
Organization: University of Oklahoma - University Computing Services

In article <1992Sep11.171500.1@hmivax.humgen.upenn.edu>, dreyfuss@hmivax.humgen.upenn.edu writes...
>Hello--
> 
>Does anyone out there know about the composition of TYE medium for growing
>E.coli TG1 cells?
> 
>Thanks,
>Jeong-Kook Kim
>Howard Hughes Medical Institute/University of Pennsylvania
>Internet:  dreyfuss@hmivax.humgen.upenn.edu
>(215)898-0172

Hi,
	The following is what we use for TYE plates and TYE medium:

TYE plates:  
       10 g. of Bacto-tryptone, 5 g. of yeast extract, 8 g. of NaCl,
       15 g. of nutrient agar, distilled water to 1 liter; autoclave
       to sterilize.
       Cool to 55 deg C, add antibiotic if desired, pour into sterile
       petri dishes.  (approx. 20 ml/plate)

TYE medium:
       Same as TYE plates but without any agar.

  - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -
 \  Bruce A. Roe                     Dept. Chemistry and Biochemistry /
 /  BROE@aardvark.ucs.uoknor.edu     University of Oklahoma           \
  - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - -