From owner-biotechniques@net.bio.net Wed Apr 02 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Rong Wang Newsgroups: bionet.journals.letters.biotechniques Subject: suicide vectors Date: 3 Apr 1997 05:17:56 -0800 Organization: The University of Montana Lines: 5 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <3342C96D.1614@selway.umt.edu> NNTP-Posting-Host: net.bio.net I am looking for suicide vectors for E. coli. Any information will be appreciated. Angelika Longacre longacre@selway.umt.edu From owner-biotechniques@net.bio.net Tue Apr 08 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Peter Cherepanov Newsgroups: bionet.journals.letters.biotechniques Subject: re: suicide vectors Date: 9 Apr 1997 05:26:34 -0700 Organization: REGA Inst. for Med. Res.... Lines: 8 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <3349346E.4DD8665B@uz.kuleuven.ac.be> NNTP-Posting-Host: net.bio.net choice depends on what you need. you may consider pKNG101, Gene,109,(1991),137-141. available from Phabagen Collection (Netherlands) #PC-V3314. regards, Peter. From owner-biotechniques@net.bio.net Tue Apr 08 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Thomas Hanselmann Newsgroups: bionet.journals.letters.biotechniques Subject: Who knows papers about protoplast culture of bamboo? Date: 9 Apr 1997 05:28:52 -0700 Organization: - Lines: 8 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <01bc440f$3e11bf80$a94a87ca@pc-yin-s> NNTP-Posting-Host: net.bio.net I am a Thai student looking for papers about protoplast culture of bamboo. Any information is welcome. Please contact me through email (g37ypk@ku.ac.th). Until the end of April 97 you can contact me via hanselmann@ibm.net. Thank you very much. Yuphin From owner-biotechniques@net.bio.net Thu Apr 10 23:00:00 1997 Path: biosci!biosci!not-for-mail From: yjc-dce@mail.tsinghua.edu.cn (Dr. Yi-Feng Shi) Newsgroups: bionet.journals.letters.biotechniques Subject: Trehalose Date: 11 Apr 1997 06:15:26 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 14 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <9704110032.AA05370@mail> NNTP-Posting-Host: net.bio.net ASKING FOR HELP We have got a sample of trehalose which may contain some lactose , glucose , fructose and maltose . But now we have not got a good analysis method to determine the chemical composition of the sample . If we are told of the method (the easier,the better) or even part of it , you would be very appreciated. From owner-biotechniques@net.bio.net Thu Apr 17 23:00:00 1997 Path: biosci!biosci!not-for-mail From: "D. Hesse" Newsgroups: bionet.journals.letters.biotechniques Subject: Problems with Asp-N Date: 18 Apr 1997 06:28:30 -0700 Organization: MPI f|r experimentelle Medizin Lines: 19 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <3357661F.4F5B@exmdi1.mpiem.gwdg.de> NNTP-Posting-Host: net.bio.net I tried to cleave a small protein ( 6,7 kD )with endoproteinase asp-N in 50 mM sodium phosphate pH 8,0 + 10% acetonitril at 370C overnight. Due to amino acid analysis we are sure, that there are at leat 3 aspartic acid residues in the molecule. To check if the asp-N works, I also performed a cleavage of myoglobin under the same conditions. No fragmentation occured ! What is wrong, what should be changed ? Does anyone know a special procedure ( pretreatment of the protein )? Thank's for help ! -- Doerte Hesse Max-Planck-Institut f|r experimentelle Medizin Abt. Immunchemie Hermann-Rein-Strasse 3 37075 Goettingen Tel.: 0551 / 3899-304 Fax : 0551 / 3899-323 E-mail : Hesse@exmdi1.mpiem.gwdg.de From owner-biotechniques@net.bio.net Thu Apr 17 23:00:00 1997 Path: biosci!biosci!not-for-mail From: MMCCARTHY@biotechnet.com Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques 22(5), May 1997 Date: 18 Apr 1997 08:10:47 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 203 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <01IHUK9GSRMQ9EFL1M@delphi.com> NNTP-Posting-Host: net.bio.net BioTechniques 22(5), May 1997 Table of Contents BENCHMARKS In vivo labeling of over-expressed recombinant proteins in E. coli Giovane, C., Schwalbach, G., and Weiss,E. Expression of the highly toxic centromere binding protein CENP-B in E. coli using the pET system in the absence of the inducer IPTG.=20 Bocanegra, J.A., Bejarano, L.A., and Valdivia, M.M. Increased efficiency in screening large numbers of cDNA fragments generated by differential display Corton, J.C. and Gustafsson, J.-A. XcmI-containing vector for direct cloning of PCR products Borovkov, A.Y. and Rivkin, M.I. High efficiency human B-cell cloning using hygromycin B-resistant feeder cells Fabb, S.A. and Ragoussis, J. Rapid mapping and subcloning of genomic clones in bacteriophage lambda by PCR Woo, H.-H. and Hawes, M.C. PCR-based assay to determine nuclear matrix association Kramer, J.A. and Krawetz, S.A. Spatial visualization of apoptosis using a whole-mount in situ DNA end-labeling technique. Smith, S.M. and Cartwright, M.M. Direct in situ end labeling for detection of apoptotic cells in tissue sections Zhang, Z. and Galileo, D.S. Optimized factor V gene mutation detection using buffy-coat direct PCR Pernod, G., Mossuz, P., and Polack, B. Spectrophotometric determination of oxidative metabolism.=20 Lokuta, M.A., Mehring, G.H., and Paulnock, D.M. Protein staining with Ponceau S during acid-urea-triton-polyacrylamide gel electrophoresis Correia, H., Rivero, J., and Herrera, F. Isolation of nuclear proteins from human brains Zhang, X., Huang, C.J., Nazarian, R., Ritchie, T., de Vellis, J.S., and Noble, E.P. Use of a single sequencing termination reaction to distinguish between cytosine and 5-methylcytosine in bisulfite-modified DNA Yuanxiang, Z., Magill, J.M., Newton, R.J., and Magill, C. Hyaluronidase generates a single-cell suspension from cultured mouse lung epithelial cells. Tolwinski, N.S. and Kano-Sueoka, T. pPE1000: a versatile vector for the expression of epitope-tagged foreign proteins in transgenic plants Hancock, K.R., Phillips, L.D., White, D.W.R., and Ealing, P.M. Two products for siliconizing glass plates used in gel electrophoresis Elsom, B.L. and Herzog, N.K. Rapid method for preparing adenovirus DNA Deryckere, F. and Burgert, H.-G. Microplate digestion of minipreps from a single microcentrifuge tube Brady, J.L. and Lew, A.M. Isolation of genomic DNA from claw clippings for genetic analysis by PCR Drummond, P.B., Song, Y., Vaughn, D., Gbadamosi, A., Butler, K., and Smith, E.J. Increased yield of plasmid DNA during removal of CsCl by ethanol precipitation Hildeman, D.A. and Muller, D. Determining the potentiative state of a chromatin domain Kramer, J.A. and S.A. Krawetz THE INTERNET ON-RAMP -Cyberspace for Biologists SHORT TECHNICAL REPORTS PCR-amplified cDNA probes for verification of differentially expressed genes Ross, R., Kumpf, K., and Reske-Kunz, A.B. Agglutination-inhibition assay for the detection of recombinant proteins tagged with peptideepitopes Dolezal, O., Coia, G., and Hudson, P.J. Transient selection during vaccinia virus recombination with insertion vectors without selectable markers.=20 Kurilla, M.G. Cloning DNA fragments between two adjacent/overlapping restriction sites using a "positive stuffer" Loukianov, E., Loukianova, T., and Periasamy, M. Making hybrids of two-hybrid systems Dagher, M.-C. and Filhol-Cochet, O. Adaptation of a =E1-1,3-glucanase assay to microplate format Zheng, Y. and Wozniak, C.A. Association of enzyme inhibition with methods of museum skin preparation Hall, L.M., Willcox, M.S., and Jones, D.S. Detection of mycoplasma infection of mammalian cells Xia, H., Fitzgerald, J., Bredt, D.S., and Forsayeth, J.R. RESEARCH REPORTS An iterative and regenerative method for DNA sequencing Jones, D.H. Alternative system for detection and mapping of activation domains Askovic, S. and Baumann, R. Optimization of cellular ELISA for assay of surface antigens on human synoviocytes Smith, D.D., Cohick, C.B., and Lindsley, H.B. Microplate assay for measurement of histamine release from mast cells Price, J.A. Simultaneous detection of multiple point mutations using fluorescence-coupled competitive primer extension Fauser, S. and Wissinger, B. Collagen-coated barium-alginate microcarriers for the culture of anchorage-dependent mammalian cells Grohn, P., Klock, G., and Zimmermann, U. PRODUCT APPLICATION FOCUS Agarose-based system for separation of short tandem repeat loci White, H.W. and N. Kusukawa The PerFect Lipid Optimizer Kit for maximizing lipid-mediated transfection of eukaryotic cells Griffiths, T., Russell, M., Froning, K., Brown, B.D., Scanlon,S.M., Almazan, M., Marcil, R., and Hoeffler, J.P. NEW PRODUCTS INDEX TO ADVERTISERS From owner-biotechniques@net.bio.net Sat Apr 19 23:00:00 1997 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.journals.letters.biotechniques Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 20 Apr 1997 07:37:35 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 239 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <199704200900.CAA25378@net.bio.net> NNTP-Posting-Host: net.bio.net (LAST REVISION: 30-JUL-95) This BIOSCI "miniFAQ" is designed to answer the questions that come up the *most frequently*. The main BIOSCI FAQ (Frequently Asked Questions) is accessible on the World Wide Web at URL http://www.bio.net/. If you can not find an answer to your question in this or other documentation, the BIOSCI technical support staff answers e-mail queries sent to biosci-help@net.bio.net We can only answer questions about the use of the newsgroups and mailing lists. We unfortunately do not have the staff to do Internet information searches or answer scientific questions. Please post those to the appropriate BIOSCI/bionet newsgroups. Contents: -------- 0) BIOSCI NEEDS YOUR SUPPORT!! 1) Using the WWW to access the BIOSCI/bionet newsgroups. 2) What to do about "spams," i.e., junk mail, ads, etc. 3) Examples of subscribing and unsubscribing to the mailing lists. 4) The BIOSCI user address and research interest directory. 0) BIOSCI NEEDS YOUR SUPPORT!! ------------------------------ BIOSCI's government funding has been expended, and we are now operating solely from advertising revenue that we have raised from our Web site at http://www.bio.net/. We need just a few minutes of your time to help us serve you. You can do two important things which will take very little time for you individually and will immensely help us continue to help you. First, please use our WWW system at http://www.bio.net/ to access the archives. You can post or reply to messages via your Web browser as described in item #1 below. Your usage helps attract sponsors. If you contact any of our sponsors, please be sure to thank them for supporting BIOSCI. It is critical for them to get this feedback if they are to continue their sponsorship for the long term. Second, if you work for a company or organization that provides products or services of interest to the biology community, please pass this message on to your marketing or marketing communications department or other appropriate group. Please ask them to help support BIOSCI by sponsoring our Web site and explain the uses and benefits of the system to the biology community. If they are interested, they can then contact us for further information at our tech support address, biosci-help@net.bio.net. 1) Using the WWW to access the BIOSCI/bionet newsgroups. -------------------------------------------------------- As of 10 December 1995, all BIOSCI/bionet full newsgroups are accessible through the World Wide Web (WWW) at URL http://www.bio.net. One can read and reply publicly or privately to both recent postings and archived messages through one's Web browser if it is configured properly to send e-mail. Each newsgroup is equipped with its own WAIS index. The main BIOSCI home page also has access to the BIO-JOURNALS Table of Contents database WAIS index and the BIOSCI user address database described in another item further below. 2) What to do about "spams," i.e., junk mail, ads, etc. ------------------------------------------------------- BIOSCI is a set of parallel USENET newsgroups (the "bionet" groups), mailing lists, and a hypermail archive at URL http://www.bio.net/. The same postings are distributed on all media (except for a small number of mailing-list-only groups at net.bio.net). Unfortunately it is becoming a despicable practice on the Internet (by a few people out to make a fast buck) to do automated mass postings to thousands of newsgroups and mailing lists. These attempts to grab free advertising are refered to as "spams" in the usual, somewhat boneheaded, net terminology. USENET is more susceptible to this practice, and many spams originate on the USENET groups and then are passed on to the mailing lists. However, spammers also get lists of mailing addresses and hit these too, so neither medium is immune. What should you do personally if you get junk mail? --------------------------------------------------- Just delete it and move on without reading it further. Filing a protest is becoming increasingly useless because spammers are often disguising the addresses where the messages are sent from. Unless you really understand Internet mail systems, your attempt at protest by sending replies to the message will often end up being sent to the address of an innocent person that the spammer is victimizing. What can BIOSCI/bionet do to protect its newsgroups? ---------------------------------------------------- The only solution currently available is to moderate the newsgroup. If this newsgroup is already moderated, then you are in good shape. Moderation protects the USENET distribution from about 95% of the spams that are being sent to date and protects the mailing lists completely. Moderation means, however, that someone has to take the time to review each message before it goes out. We have set up software here that simply allows the moderator to forward to an address at net.bio.net messages that (s)he wishes to have distributed. This takes no more time than that needed to read the message and pass it on, say about 1 min. per message. Most newsgroups currently have a discussion leader who is responsible for their newsgroup. The discussions leaders and their e-mail addresses are listed in the BIOSCI Information Sheet which is available on the Web at http://www.bio.net/. If a newsgroup is being hit with too many junk postings, please contact the discussion leader for that group and see if there is interest in moderating the group. Please do not assume that by simply posting a complaint to the newsgroup itself, anyone on the BIOSCI staff will act on your complaint. With close to 100 newsgroups to run, the BIOSCI staff has to rely on the discussion leaders of each newsgroup to report problems directly to us at biosci-help@net.bio.net. We will moderate any of our newsgroups if the discussion leader tells us that the readership of the group wishes to do so and if a moderator is willing to do the work. For most BIOSCI/bionet groups, this entails only a few minutes of work each day. Moderating a newsgroup will resolve probably 95% of the junk postings on the USENET distribution. Unfortunately there are easy ways for determined spammers to override the moderation mechanism on USENET, but we can protect our e-mail subscribers from unwanted postings if the newsgroup is moderated. You can also access our newsgroups over the WWW at URL http://www.bio.net. While this Web interface will not stop spammers from trying to post to the groups, this will give you yet another way, besides using USENET news, to keep the junk out of your personal mail files. For those of you with local USENET news systems, the Web interface will also give you faster access to new newsgroups and recent postings. 3) Examples of subscribing and unsubscribing to the mailing lists. ------------------------------------------------------------------ PLEASE NOTE: The BIOSCI management does NOT act on subscription/unsubscription requests that are posted improperly to the newsgroups and mailing lists. People who do this only bother everyone on the lists to no avail. Please be sure to follow the proper procedures below. Gory details are in the BIOSCI Information sheets on the Web at http://www.bio.net. Below we give an example utilizing the METHODS-AND-REAGENTS list at both of our two BIOSCI sites: Users in the Americas and Pacific Rim countries who use the BIOSCI ------------------------------------------------------------------ node at computer net.bio.net: ---------------------------- A) Determine the "listname" which is the <=8 character mail address ^^^^^^^^^^^^^ for the group. These can be found in the BIOSCI Info. Sheet. For the METHODS-AND-REAGENTS group the mailing address is methods@net.bio.net. The listname is the portion of the address to the left of the @ sign, i.e., "methods". The listname is used with the "subscribe" and "unsubscribe" commands illustrated below. B) Mail all commands in the body of a mail message addressed to biosci-server@net.bio.net. Do NOT send commands to the newsgroup posting addresses! Leave the Subject: line blank, any text on it will be ignored. C) In the body of your message put one or more of the following commands with an "end" command on the last line, e.g., subscribe methods unsubscribe methods end Do NOT put your e-mail address or other text on these lines. The server only allows you to cancel your subscription if the address on your mail header matches the address on our mailing list. Please ask for help at biosci-help@net.bio.net if your address has changed, e.g., if you know you are on the list but the server tells you that you are not a member. Users in Europe, Africa, and Central Asia who use the BIOSCI node at -------------------------------------------------------------------- computer daresbury.ac.uk (also known as dl.ac.uk): ------------------------------------------------- To subscribe and unsubscribe to/from the BIOSCI lists, you need to specify the full USENET newsgroup name with "bionet-news." prepended. The USENET newsgroup names are listed in the BIOSCI Information sheet on the Web at http://www.bio.net/. For the METHODS-AND-REAGENTS list the USENET newsgroup name is bionet.molbio.methds-reagnts, thus the appropriate commands are sub bionet-news.bionet.molbio.methds-reagnts unsub bionet-news.bionet.molbio.methds-reagnts These commands are included in a message addressed to mxt@dl.ac.uk, NOT to the newsgroup mailing addresses. As usual, include the text in the body of the message as text on the Subject: line is ignored. To unsubscribe from all the lists at the UK node, use unsub bionet-news Please note that if the address in the list is different than the one in your mail message header, you will not be able to unsubscribe by this method. If you have problems, please mail biosci@daresbury.ac.uk. 4) The BIOSCI user address and research interest directory. ----------------------------------------------------------- Please take this opportunity to add your name, address, and research interest information to the BIOSCI User Address Database if you have not already done so. You can fill out the address form directly through our Web page at URL http://www.bio.net/adrform.html. The address database is reindexed nightly for WWW access (the URL is http://www.bio.net/). If you are not directly on the Internet but can reach it by e-mail, please use our waismail server to access the user directory. waismail use is described above. You can also request a user address form by e-mail from biosci-help@net.bio.net. Please check your database entry from time-to-time to see if your address information is still up-to-date. Because of our limited personnel resources, we ask that you resubmit a *complete* form to revise your entry; we only replace complete entries and do not have resources to edit old forms. Sincerely, Dave Kristofferson BIOSCI/bionet Manager biosci-help@net.bio.net From owner-biotechniques@net.bio.net Sun Apr 27 23:00:00 1997 Path: biosci!biosci!not-for-mail From: William Reisdorf Newsgroups: bionet.journals.letters.biotechniques Subject: Pichia pastoris expression systems Date: 28 Apr 1997 05:38:53 -0700 Organization: Washington University School of Medicine Lines: 15 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <3362DEB5.64C3@ibc.wustl.edu> NNTP-Posting-Host: net.bio.net > > Our lab is considering using the Zeocin-TM Pichia expression system > from Invitrogen and we're looking for any helpful comments from > others who may have used this particular system. Positive and negative > experiences would be appreciated. Our goal is to express IgSF domains > from a receptor tyrosine kinase, and so far our luck in refolding > them from E. coli inclusion bodies has been poor. We hope that the > disulfides will be able to form better and that glycosylation may > also help with the stability of our recombinant protein. > > William Reisdorf (postdoc) > Washington University > Biochem & Molec Biophys and > Inst for Biomed Computing > reisdorf@ibc.wustl.edu From owner-biotechniques@net.bio.net Mon Apr 28 23:00:00 1997 Path: biosci!biosci!not-for-mail From: Robyn Puffenbarger Newsgroups: bionet.journals.letters.biotechniques Subject: protoplast buffer and lysis buffer Date: 29 Apr 1997 06:18:14 -0700 Organization: Medical College of Virginia/VCU Lines: 13 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <3365E158.5F4A@gems.vcu.edu> NNTP-Posting-Host: net.bio.net April 29, 1997 Hello, I have the protocol for the "in-well-lysis" procedure, but have lost the recipies for the protoplast buffer and the lysis buffer. This simple technique lets you quickly see the difference in recombinant plasmids without a 'miniprep.' If anyone has these two recipies and can post them to me or to the list, I would appreciate it. Thanks in advance, Robyn Puffenbarger