HUM-MOLGEN archive: BIOT: various

The BIOTechnology/molecular biology section is open for request or offering of technical help; new protocols, problems with apparatus, exchange of experiences with protocols, apparatus etc; but also for funds offering and requests etc. Semi-commercial messages in the interest of our subscribers are allowed. Please respond by private E-mail unless your response is of general interest to all HUM-MOLGEN subscribers Thank you for your numerous contributions with our previous BIOT message! The editors Martin Kennedy Arthur Bergen *********************************************************************** Subj: BIOT mars@bioch.ox.ac.uk (Raoul Heller) sent the following comments: I would like to identify a single hamster chromosome in a monochromosomal hybrid cell line of human origin. Attempts to paint the hamster chromosome specifically using labelled total hamster genomic DNA with Cot 1 DNA weren't particularly successful: the obtained signal after FISH was weak and there was crosshybridisation. Another approach might be to amplify species specific interspersed- repetitive-element (IRE) and inter-IRE sequences which could then be labelled as probe for chromosome painting. The protocoll by David J. Munroe et al., IRE-Bubble PCR: A rapid method for efficient and representative amplification of human genomic DNA sequences from complex sources. (Genomics 19:506-514) sounds attractive in this context. Could anybody with experience in this field give me some advice on a) the practicability of this and related PCR methods if applied to generate murine or other vertebrate specific chromosome paints; b) where to find descriptions of useful IREs in various mammalian/ vertebrate species (hamster, mouse, frog, avian etc.); c) whether other strategies for the diagnosis of a single chromosome or chromosome fragment against a genetic background of a different species are more reliable in his/her practical experience? Thanks very much in advance for any helpful comments, Raoul Heller CRC Chromosome Molecular Biology Group Dept. of Biochemistry Univ. of Oxford // U.K. OX1 3QU phone: +44-1865-275225 fax : +44-1865-275259 email: mars@bioch.ox.ac.uk ************************************************************************** This message was originally submitted by EIBEL@NZ11.UKL.UNI-FREIBURG.DE to the HUM-MOLGEN list at NIC.SURFNET.NL. The video camera from our gel documentation video system from CYBERTECH (Berlin, Germany) broke recently. We tried to contact Cybertech repeatedly but we got no response except for the message from a tape recorder. Therefore we do not know whether Cybertech still exiist or how to get in touch with them. In the past, we also had a really bad impression from their coustomer service. - Did anybody make the same experience with Cybertech ? - Does anybody known how to deal with this problem ? Dr. Hermann Eibel Klinische Forschergruppe f. Rheumatologie Klinikum der Universitaet Freiburg Breisacher Str. 64 D-79106 Freiburg, Germany Tel.: 49 - 761 - 270 - 5294 Fax: 49 - 761 - 270 - 5298 ************************************************************************** Subj: BIOT: Digital Imagers-Image Analysis for Science This message was originally submitted by mmolenda@STUDENTS.WISC.EDU to the HUM-MOLGEN list at NIC.SURFNET.NL. We at Digital Imagers recently merged with BioImaging Technologies and now carry Image Analysis Systems/Software to go along with the microplate readers/washers (from Tecan/SLT) and our Ambis Image Analysis Systems. The Image Analysis Systems are either for Mac or PC. The Mac systems utilize the world renowned NIH Image to get you the best possible data and to keep your costs to a minimum. Why pay $2000 for a piece of image analysis software when you can have NIH Image for $0! Also, get some of the best technical support in the field today. With over 5 years of experience, we were there at the start of the image analysis boom, and we still remain a mainstay. Contact us today at: e-mail mmolenda@students.wisc.edu or call 608-243-9706 ************************************************************************* Subj: A technical problem for help This message was originally submitted by gpma66@UDCF.GLA.AC.UK to the HUM-MOLGEN list at NIC.SURFNET.NL. I have difficulty in transfecting constructs into either human myelomonocytic cell line ( HL-60) or mouse myelomonocytic cell line ( 32D Cl3). I have tried a wide range of electroporation conditions with the Easyject apparatus made by EquiBio from Belgium in conjunction with the luciferase assay and beta-gal assays without any success. In addition to the signle pulse, the double pulses approach has also been tried. Although this problem has been informed to the manufacturer several months ago, I have not got any feedback so far. Any suggestion and help will be highly appreaciated. Jingde Zhu CRC Department of Medical Oncology Glasgow University Gasrcube Estate, Switchback Road, Glasgow G61 1BD. U. K., ************************************************************************* From: IN%"ED-MOLGEN@nic.SURFnet.nl" "Human Molecular Genetics Editors" 20-NOV-1995 16:42:10.87 To: IN%"ED-MOLGEN@nic.SURFnet.nl" "Multiple recipients of list ED-MOLGEN" CC: Subj: DNA problems This message was originally submitted by 103202.1502@COMPUSERVE.COM to the HUM-MOLGEN list at NIC.SURFNET.NL. Hello! I need help. I'm working in cancer genetics and have been experiencing problems with the DNA that I have extracted. While most of samples work there are a few (from patients who have recently or who are receiving chemo- or radiation therapy) which don't seem to work under my normal conditions. I believe the DNA is fragile and starting to splinter. I was wondering if anyone had any ideas on what to do...ie add a conditioning reagent to my PCR buffer, etc... Thank you. deAnne Guarino/USA ****************************************************************************