From owner-biotechniques@net.bio.net Mon Mar 09 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Mary McCarthy Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques 24(3), March 1998 Date: 10 Mar 1998 08:39:21 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 143 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6e3qbp$5pt@net.bio.net> NNTP-Posting-Host: net.bio.net BioTechniques 24(3), March 1998 Tale of Contents Benchmarks Low-Voltage Separation of Phosphoamino Acids by Silica Gel=20 Thin-Layer Electrophoresis in a DNA Electrophoresis Cell S.C. Hardin and S.M. Wolniak In Situ Detection Method for Glutaminyl Cyclase Activity in Polyacrylamide Gels J.B. Houseknecht, J.S. Temple and R.C. Bateman, Jr. Analysis of Cell-Cycle Profiles in Transfected Cells Using a=20 Membrane-Targeted GFP W. Jiang and T. Hunter Chemiluminescence-Based Detection of Minute Amounts of=20 Apoptotic DNA F.Lopez Blanco, J. Gonzalez-Reyes, L.F. Fanjul, C.M. Ruiz de Galarreta and J. Quintana Aguiar Rapid Screening of Fusion Protein Recombinants by Measuring=20 Effects of Protein Overexpression on Cell Growth G.D. Davis and R.G. Harrison Versatile Low-Copy-Number Plasmids for Temperature-Inducible=20 Overexpression of Bacterial Genes in Escherichia coli C. Gotting, G. Thierbach, A. Puhler and J. Kalinowski =20 Chemiluminescence-Based RNase Protection Assays for Simultaneous Quantification of Procollagen mRNAs Containing AU-Rich Regions M.J. Ausserlechner, R. Sgonc and G. Wick Modification of Primer Design Facilitates the Use of Differential Display M.S. Brenz Verca, S. Brenz Verca, S. Rusconi and J.-L. Dreyer Use of Novel Downstream Primers for Differential Display RT-PCR X. Wang, X. Li and G.Z. Feuerstein Rapid Cloning of Telomere-Associated Sequence Using Primer- Tagged Amplification G. Fu and D.C. Barker PCR-Assisted cDNA Cloning: A Guided Tour of the Minefield R. Hooft van Huijsduijnen Use of the Restriction Enzyme AvaI and Exo- Bst Polymerase in Strand Displacement Amplification M.A. Milla, P.A. Spears, R.E. Pearson and G.T. Walker =20 Long-Distance Genome Walking Using the Long and Accurate=20 Polymerase Chain Reaction G.-S. Min and J.R. Powell Relative Amplification Efficiency of Differently Sized Templates by Long-Distance PCR G. Sanchez, D. Gautheret, X. Xu, A.-L. Chenine and I. Hirsch Re-amplification of Short Primer-Generated Bands from RAPD and Methylation-Sensitive Restriction Fingerprinting by Discrimination Primers M. Zeng, E.O. Martsen and J.-N. Lapeyre Direct Amplification of Microsatellite Alleles from Sonicated=20 Goldfish Sperm W. Zheng, N. Stacey, D. Liu and C. Strobeck Construction of a Device Composed of Common Plumbing Supplies=20 for Freezing Microscopy Samples D.W. Darnowski and L.O. Vodkin Simultaneous Purification of RNA and DNA from Liver Using=20 Sodium Acetate Precipitation J.K. Evans, P. Troilo and B.J. Ledwith Internet On-Ramp Cyberspace for Biologists Short Technical Reports Random Mutagenesis by Whole-Plasmid PCR Amplification A. Parikh and F.P. Guengerich Long PCRs of Transposons in the Structural Analysis of Genes=20 Encoding Acquired Glycopeptide Resistance in Enterococci H. Haaheim, K.H. Dahl, G.S. Simonsen, O. Olsvik and A. Sundsfjord Bacterial Growth Medium that Significantly Increases the Yield of Recombinant Plasmid H.M. Duttweiler and D.S. Gross 5=A2-Degenerate 3=A2-Dideoxy-Terminated Competitors of PCR Primers Increase Specificity of Amplification S.P. Atamas, I.G. Luzina, B.S. Handwerger and B. White=20 Small-Scale Isolation of Genomic DNA from Streptomyces Mycelia=20 or Spores A.J. Kutchma, M.A. Roberts, D.B. Knaebel and D.L. Crawford =20 Simultaneous Detection of Two GFP Spectral Mutants During In Vivo Confocal Microscopy of Migrating Dictyostelium Cells T. Zimmerman and F. Siegert Research Reports Development and Applications of Enhanced Green Fluorescent=20 Protein Mutants R.H. Stauber, K. Horie, P. Carney, E.A. Hudson, N.I. Tarasova, G.A. Gaitanaris and G.N. Pavlakis Localization of Trinucleotide Repeat Sequences in Myotonic Dystrophy Cells Using a Single Fluorochrome-Labeled PNA Probe K.L. Taneja Product Application Focus Fusion of Green Fluorescent Protein with the Zeocin=D4-Resistance=20 Marker Allows Visual Screening and Drug Selection of Transfected Eukaryotic Cells R.P. Bennett, C.A. Cox and J.P. Hoeffler FastTag Nucleic Acid Labeling System: A Versatile Method for Incorporating Haptens, Fluorochromes and Affinity Ligands into=20 DNA, RNA and Oligonucleotides S.G. Daniel, M.E. Westling, M.S. Moss and B.D. Kanagy Ad Hoc Reviewers Literature Express New Products Index to Advertisers From owner-biotechniques@net.bio.net Wed Mar 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Ezzat El-Akkad Newsgroups: bionet.journals.letters.biotechniques Subject: BioCitation Date: 19 Mar 1998 12:13:37 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 6 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6eru9h$noj@net.bio.net> Reply-To: ezzatak@ultinet.net NNTP-Posting-Host: net.bio.net Eaton Publishing Co. 154 E. Central St. Natick, MA 01760-9898 USA Tel: 508-655-8282 FAX: 508-655-4356 From owner-biotechniques@net.bio.net Wed Mar 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Lolis, Elias" Newsgroups: bionet.journals.letters.biotechniques Subject: mutagenesis strategy Date: 19 Mar 1998 13:09:24 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 18 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6es1i4$uu@net.bio.net> NNTP-Posting-Host: net.bio.net I would like to randomly mutate all 7 transmembrane regions of G protein-coupled receptors and then select for mutated receptors with functional properties. I was wondering whether anyone knows of any method that could target different regions of a cDNA for simultaneous random mutagenesis. One strategy I thought of requires a mutagenesis method that selectively targets either single-stranded or double-stranded DNA. For example, if there is a method that selectively targets single-stranded DNA, I could produce a single-stranded version of the cDNA, protect the non-transmembrane regions by synthesizing complementary DNA for these sequences, and applying the mutagenesis procedure to the remaining single-stranded DNA. If anyone knows of a method or article that answers this question please send me e-mail. Thanks. Elias Lolis From owner-biotechniques@net.bio.net Wed Mar 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Ezzat El-Akkad Newsgroups: bionet.journals.letters.biotechniques Subject: BioCitation Date: 19 Mar 1998 12:33:24 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 31 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6ervek$q9f@net.bio.net> Reply-To: ezzatak@ultinet.net NNTP-Posting-Host: net.bio.net There is an article referenced in the latest Internet On-Ramp (BioTechniques, Vol. 24, No. 3, March 1998) which appeared in BioTechniques back in 1994. We do not have any issues which go back that far. Can anyone supply us with the article? J. Tesgaigzi, W. Smith-Harrison and D.M. Carlson, 1994, A simple method for reusing Western blots on PVDF membranes, BioTechniques 17:268-269. In addition, can anybody supply a similar procedure for Nitrocellulose membranes? Please respond to: WDSPIV@UNFORGETTABLE.COM Mary McCarthy, PhD Associate Scientific Editor BioTechniques Eaton Publishing Co. 154 E. Central St. Natick, MA 01760-9898 USA Tel: 508-655-8282 FAX: 508-655-4356 From owner-biotechniques@net.bio.net Wed Mar 18 22:00:00 1998 Path: biosci!biosci!not-for-mail From: Mary McCarthy Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques 24(4), April 1998 Date: 19 Mar 1998 13:21:35 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 133 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6es28v$2na@net.bio.net> NNTP-Posting-Host: net.bio.net BioTechniques 24(4), April 1998 Table of Contents Benchmarks Double Labeling with Fluorescence In Situ Hybridization in Drosophila Whole-Mount Embryos S.C. Hughes and H.M. Krause Fluorescence In Situ Hybridization of Cells on Polyvinylidene Fluoride Membrane Filters A. Mahr and A.E.Bowe Standardizing the Counting of Adipocytes in Cell Culture K. Gorzelniak, S. Engeli and A.M. Sharma Plasmid Maintenance Assay Based on Green Fluorescent Protein and FACS of Mammalian Cells M. Vogel, K. Wittmann, E. Endl, G. Glaser, R. Knuchel, H. Wolf and H.H. Niller Fluorescent Escherichia coli C for Enumeration of Coliphages from Environmental Samples N. Jothikumar and D.O. Cliver Screening for Recombinant E. coli Using Antibiotic Test Strips U. Reischl, C. Gerdes and I.P. Thrippleton Detergent and Enzyme Treatment of Apoptotic Cells for the Observation of DNA Fragmentation D.J. Park and P.Q. Patek Construction of a Genomic DNA Library by TA Cloning Y. Kawata, S. Yano and H. Kojima Generation of Genomic Mini-Libraries by Taq DNA Polymerase Modification of Genomic Fragments H.-J. Huang and C. Pears Small-Scale Preparation of the Single-Copy Bacterial Artificial Chromosome Vector pBeloBAC11 D. Tillett and B.A. Neilan PCR Amplification of cDNA Libraries for Cloning and Screening T.H. You and R.L. Scholl 3-prime RACE: Skewed Ratio of Specific to General PCR Primers Improves Yield and Specificity I.N. Bespalova, S. Adkins and M. Burmeister Distinct Combination of Purification Methods Dramatically Improves Cohesive-End Subcloning of PCR Products W.A. Wybranietz and U. Lauer Avoiding False Positives in Colony PCR Q. Dallas-Yang, G. Jiang and F.M. Sladek Digestion of Terminal Restriction Endonuclease Recognition Sites on PCR Products K. Zimmermann, D. Schogl and J.W. Mannhalter Pitfalls of Processed Pseudogenes in RT-PCR H. Mutimer, N. Deacon, S. Crowe and S. Sonza High-Voltage and High-Salt Buffer Facilitates Electroporation of Human Aortic Smooth-Muscle Cells C.-N. Yan, F. Li, C. Patterson and M.S. Runge Efficient Cloning of DAF Polymorphic Markers from Silver-Stained Polyacrylamide Gels A.E. Men and P.M. Gresshoff Efficient Large-Scale Transformation of Yeast K. Yamada, J.-C. Wang, H. Osawa, D.K. Scott and D.K. Granner BioFeedback Internet On-Ramp Cyberspace for Biologists Short Technical Reports Herpes Simplex Virus Thymidine Kinase-Green Fluorescent Protein Fusion Gene: New Tool for Gene Transfer Studies and Gene Therapy S. Loimas, J. Wahlfors and J. Janne Quantitative RT-PCR Amplification of RNA in Single Mouse Oocytes and Preimplantation Embryos M.T. Fiorenza and F. Mangia Lac/Tet Dual-Inducible System Functions in Mammalian Cell Lines H.-S. Liu, C.-H. Lee, C.-F. Lee, I.-J. Su and T.-Y. Chang Establishment of a Whole-Chick Sternum Model that Recapitulates Normal Cartilage Development M.S. Hirsch and K.K.H. Svoboda Vectors to Target Protein Domains to Different Cellular Compartments G. Toby, S.F. Law and E.A. Golemis Research Reports Intracellular Magnetic Labeling of Lymphocytes for In Vivo Trafficking Studies U. Schoepf, E.M. Marecos, R.J. Melder, R.K. Jain and R. Weissleder Reproducibility in the Quantification of mRNA Levels by RT-PCR-ELISA and RT Competitive-PCR-ELISA L.L. Hall, G.R. Bicknell, L. Primrose, J.H. Pringle, J.A. Shaw and P.N. Furness Retroviral Gene Transfer in Chondrogenic Limb Bud Micromass Cultures N.S. Stott, Y.-S. Lee and C.-M. Chuong Diffusion of Proteins Across the Nuclear Envelope of HeLa Cells S. Chatterjee and U. Stochaj Product Application Focus SDS Agarose Gels for Analysis of Proteins M. Wu and N. Kusukawa New Products Index to Advertisers From owner-biotechniques@net.bio.net Thu Mar 19 22:00:00 1998 Path: biosci!biosci!not-for-mail From: BIOSCI Administrator Newsgroups: bionet.journals.letters.biotechniques Subject: BIOSCI/bionet miniFAQ & Fundraiser Date: 20 Mar 1998 06:03:16 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 236 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6etsv4$hlr@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. From owner-biotechniques@net.bio.net Thu Mar 19 22:00:00 1998 Path: biosci!biosci!not-for-mail From: bmbthc@bmb.leeds.ac.uk (Hedley Carr) Newsgroups: bionet.journals.letters.biotechniques Subject: Yeast protein isolation for Immunoblot... Date: 20 Mar 1998 06:03:16 -0800 Organization: University of Leeds, UK. Lines: 11 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6etsv4$hm0@net.bio.net> Reply-To: bmbthc@bmb.leeds.ac.uk NNTP-Posting-Host: net.bio.net Hi, I'm looking for a straight-forward method of smashing up / lysing yeast (S. cerevisiae) cells. I'm trying to perform Western blots with the lysed material but I get very strong background staining with what is normally a good antibody. I think it might be cell wall components or something but does anyone do this regularly? If so, what's your method? Thanks for you time! From owner-biotechniques@net.bio.net Mon Mar 23 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Hiroyuki Naito" Newsgroups: bionet.journals.letters.biotechniques Subject: How to the Transfection Date: 24 Mar 1998 09:01:41 -0800 Organization: SSP Lines: 16 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6f8otl$er4@net.bio.net> NNTP-Posting-Host: net.bio.net I have a trial for transient transfection to ATL-16T cell-line by the electropolation. But it is in vain. So I wish to know about the "cationic polymer" to use to transfect the DNA(plasmid) to cultured ATL-16T. Do you know the best reagent to achieve this work ? please responed to: hz6h-nitu@asahi-net.or.jp Hiroyuki Naito Narita/Chiba/Japan From owner-biotechniques@net.bio.net Mon Mar 23 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Piotr Solarz" Newsgroups: bionet.journals.letters.biotechniques Subject: Renaturation of plant acid phosphatase Date: 24 Mar 1998 09:01:41 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 25 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6f8otl$er8@net.bio.net> NNTP-Posting-Host: net.bio.net Dear collegues, I am seeking for a biochemist that has an experience in refolding denaturated plant acid phosphatases. I mean a person who could advice me what bufers, substances, temperatures... use first. Thank you. -- * * * * * * * * * * * * * * * * Piotr Solarz __________________________________________________ Uniwersytet Wroclawski Instytut Biochemii i E-mail: Biologii Molekularnej PSolarz@IC.COM.PL ul. Tamka 2 50-137 WROCLAW ICQ UNI 4572571 POLAND _________________________________________________ From owner-biotechniques@net.bio.net Mon Mar 23 22:00:00 1998 Path: biosci!biosci!not-for-mail From: "Dion Florack" Newsgroups: bionet.journals.letters.biotechniques Subject: Re: mutagenesis strategy Date: 24 Mar 1998 09:01:45 -0800 Organization: World Online Lines: 21 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6f8otp$eri@net.bio.net> NNTP-Posting-Host: net.bio.net Dear Lolis, Elias, using the Chameleon mutagenesis kit you can mutate several regions in a single event. Most likely you can also use this kit/strategy for random mutagenesis of more than one region. Since one of the oligos that you will need removes an unique restriction site you will only target one of the two strands. The non-mutated one is removed by restriction endonuclease treatment using the enzyme for which the site was removed in the mutagenesis event. If you need more info on this protoclo, send me an email at florack@usa.net and I will contact you. Greetings, Dion Florack. Lolis, Elias heeft geschreven in bericht <6es1i4$uu@net.bio.net>... > I was wondering whether anyone knows of any method >that could target different regions of a cDNA for simultaneous random >mutagenesis. From owner-biotechniques@net.bio.net Thu Mar 26 22:00:00 1998 Path: biosci!biosci!not-for-mail From: luke_c@leila.tch.harvard.edu (Cliff Luke) Newsgroups: bionet.journals.letters.biotechniques Subject: Far Western Date: 27 Mar 1998 13:57:34 -0800 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 24 Sender: daemon@net.bio.net Approved: mmccarthy@BioTechniques.com Distribution: world Message-ID: <6fh7ce$kl4@net.bio.net> NNTP-Posting-Host: net.bio.net Hi. I am looking for as many techniques as I can find for Far Western analysis using protease inhibitors against either purified proteases or cell lysates. I have tried several different methods but so far have been unsucessful. Thanks Cliff Luke __________________ Cliff Luke luke_c@leila.tch.harvard.edu Joint Program in Neonatolgy, Childrens Hospital, Enders 970, Boston MA 02115