From owner-biotechniques@net.bio.net Wed Jun 19 23:00:00 1996 Path: biosci!biosci!not-for-mail From: "Larry H. Yamaoka" Newsgroups: bionet.journals.letters.biotechniques Subject: Chelex 100 Date: 20 Jun 1996 16:15:26 -0700 Organization: Duke University Medical Center Lines: 11 Sender: daemon@net.bio.net Approved: biosci-help@net.bio.net Distribution: world Message-ID: <31C95043.430C@dnadoc.mc.duke.edu> NNTP-Posting-Host: net.bio.net In the June, 1996 BioTechniques, Del Rio et al extract DNA from the punch using Chelex 100. Is the extraction carried out on the punch after phenol extraction or directly without previous treatment? On the extraction of DNA using Chelex, does anyone have experience using this procedure on paraffin sections--that is, would chelex extraction work on fixed, paraffin embedded tissue? Thank you for any information provided. From owner-biotechniques@net.bio.net Thu Jun 20 23:00:00 1996 Path: biosci!biosci!not-for-mail From: MMCCARTHY@biotechnet.com Newsgroups: bionet.journals.letters.biotechniques Subject: BioTechniques 21(1), July 1996 Date: 21 Jun 1996 10:15:30 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 142 Sender: daemon@net.bio.net Approved: MMCCARTHY@BIOTECHNET.COM Distribution: world Message-ID: <01I666RQMECI8YC4T7@delphi.com> NNTP-Posting-Host: net.bio.net BioTechniques 21(1), July 1996 Table of Contents BENCHMARKS Cycle sequencing protocol using DeepVent(exo-) DNA polymerase and reduced dNTP and =E0-35S-dATP concentrations -Mariam=82 B. Random primed gene walking PCR: a simple procedure to retrieve nucleotide fragments adjacent to known DNA sequences -Trueba G.A. and R.C. Johnson Improved PCR amplification of multiple specific alleles (PAMSA) using internally mismatched primers -Okimoto R. and J.B. Dodgson Salt-dependent performance variation of DNA polymerases in co-amplification PCR -Favre, N. and W. Rudin Amplification of DNA from whole cells of cyanobacteria using PCR -Howitt C.A Improved ligation-anchored PCR strategy for identification of 5' ends of transcripts -Ansari-Lari M.A., S.N. Jones, K.M. =20 Timms and R.A. Gibbs Microdissection RT-PCR analysis of gene expression in pathologically defined frozen tissue sections -Hiller T., L. Snell and P.H. Watson Dimethyl sulfoxide improves RNA amplification -Sidhu M., M.-J. Liao and A. Rashidbaigi Preparation of transcriptionally active nuclear extracts from mammalian tissues -Stuempfle K.J., M. Koptides, A.M. =20 Karinch and J. Floros Preparation of single-stranded phagemid DNA without chromosomal DNA contamination -Tang W.T., J.-N. Liu and V. Gurewich RNA/DNA mini-prep from a single sample of orchid tissue -Knapp J.E. and J.M. Chandlee Improved purification and yields of RNA by RNeasy -Bonham M.J. and D. Danielpour Monitoring nuclear transport in HeLa cells using the green fluorescent protein -Chatterjee S. and U. Stochaj Generation of recombinant baculoviruses by direct cloning -Lu A. and L.K. Miller Low cost agarose gel documentation system -Scott T.M., G.L. Dace and M. Altschuler THE INTERNET ON-RAMP Cyberspace for Biologists SHORT TECHNICAL REPORTS Application of 5-bromo-2'deoxyuridine as a label for in situ hybridization in chromosome microdissection and painting, and 3'OH DNA end labeling for apoptosis -M=81hlman-D=A1az M.C., R.G. Dullea and =20 J.S. Bedford Uncultured blood samples can be labeled by PRINS and ready for chromosome enumeration analysis 1 H after collection -Gosden J. and G. Scopes Comparison of plasmid DNA preparation methods for direct gene transfer and genetic immunization -Davis H.L., M. Schleef, P. Moritz, M. =20 Mancini, J. Schorr and R.G. Whalen Partial CviJI digestion as an alternative approach to generate cosmid sublibraries for large-scale sequencing projects -Gingrich J.C., D.M. Boehrer and S.B. =20 Basu RESEARCH REPORTS NIRCA: A rapid robust method for screening for unknown point mutations -Goldrick M.M., G.R. Kimball, Q.Liu, =20 L.A. Martin, S.S. Sommer and J.Y. Tseng Solid-phase method for differential display of genes expressed in hematopoietic stem cells -Rosok O., J. Odeberg, M. Rode, T. =20 Stokke, S. Funderud, E. Smeland and J. Lundeberg Colorimetric solid-phase capture hybridization assay for detection of amplified Borrelia burgdorferi DNA -Mansy F., B. Hoyois, M.-J. De Vos, A. =20 Van Elsen, A. Bollen and E. Godfroid Cytosine methylation: quantitation by automated genomic sequencing and Genescan analysis -Paul C.L. and S.J. Clark Experimental design: a useful tool for PCR optimization -Boleda M.D., P. Briones, J. Farr=82s, L. Tyfield and R. Pi PRODUCT APPLICATION FOCUS Agarose gel analysis of 15-40 kb PCR amplimers -Burland V., F.P. Curtis and N.Kusukawa LITERATURE EXPRESS NEW PRODUCTS RECRUITMENT INDEX TO ADVERTISERS From owner-biotechniques@net.bio.net Mon Jun 24 23:00:00 1996 Path: biosci!biosci!not-for-mail From: RHODES@UConnVM.UConn.Edu Newsgroups: bionet.journals.letters.biotechniques Subject: Fluorescence standard Date: 24 Jun 1996 18:00:04 -0700 Organization: BIOSCI International Newsgroups for Molecular Biology Lines: 26 Sender: daemon@net.bio.net Approved: biosci-help@net.bio.net Distribution: world Message-ID: <4qndmk$ps@net.bio.net> NNTP-Posting-Host: net.bio.net We're in the process of testing the sensitivity of a fluorescent probe to a variety of related macromolecules, and will be doing LOTS of standard curves in the very near future. The instrument we're using is a Spex fluorometer, which means that we have 2 sets of 2 slits that need to be set each time we do the experiments (since there are other projects that have different requirements using the machine in-between these runs). We run a background spectrum every time, so that we know what to correct, but if we're doing intensity vs. concentration over a long period of time, we'll need a stable standard to normalize the runs from day-to-day. (((( BOTTOM LINE QUESTION STARTS HERE )))) Can anyone suggest a _stable_ fluorophore that we could use as a standard? Something that will not bleach out, precipitate, etc. over the course of the next few months... Thanks lots - As usual, if there is interest or debate, I'll post the responses. | O==O | | DAVID G. RHODES O==O PHONE 860-486-5413 | | SCHOOL OF PHARMACY; U-92 O==O FAX 860-486-4998 | | UNIVERSITY OF CONNECTICUT O==O | | STORRS, CT 06269-2092 O==O RHODES@UCONNVM.UCONN.EDU | | O==O |