Cold Spring Harbor Laboratory

CLONING & ANALYSIS OF LARGE DNA MOLEULES

APRIL 8-21, 1997

This course covered the theory and practice of manipulating, cloning, and mapping large segments of DNA. The course focused on the hands-on construction of bacterial artificial chromosome (BAC) clone libraries, and the use of several types of large-insert clone libraries (YAC, BAC, P1) and genomic DNA for the analysis of large chromosome regions in the physical mapping and contig-building phases of positional cloning projects. Lectures and laboratory work included BAC clone library preparation, preparative and analytical pulsed field gel electrophoresis (PFGE), techniques for the isolation and manipulation of high molecular weight DNA from a variety of sources (including plants and animals), and an introduction to relevant bacterial and yeast genetics. BAC- and YAC-based positional cloning methods included library screening, contig assembly, long range restriction and RecA-assisted Restriction Endonuclease (RARE) cleavage mapping, and recovery/analysis of large-insert clone end-fragments. Lectures by outside speakers on topics of current interest supplemented the laboratory work.

INSTRUCTORS:

Abderrahim, Hadi, Stanford University (hadi@genome. stanford .edu)

Riethman, Harold, The Wistar Institute (Riethman@wista.wistar.upenn. edu)

Wing, Rod, Clemson University (rwing@clemson.edu)

Dewar, Ken, Whitehead, MIT (kdewar@genome.wi.mit.edu)

Choi, Sangdun, California Institute of Technology (schoi@caltech.edu)

Frisch, David, Clemson University (dfrisch@clemson.edu)

SPEAKER LIST :

Vladimir Larionov, NIH (larionov@niehs.nih.gov) : TAR cloning

Andreas Gnirke, Mercator Genetics (gnirke@mercator.com) : RARE cleavage

Martin Ganal (ganal@ipk-gatersleben.de) : Positional cloning in plants

Roger Reeves, Johns Hopkins Univ. Schl. Med. (rreeves@welchlink.welch.jhu.edu) : Comparative mouse and human genomics

Bruce Birren, WhiteHead, MIT (bwb@genome.wi.mit.edu) : Pulsed Field Gel Electrophoresis

Vivian Cheung, University of Pennsylvania (cheungv@email.chop.edu) : Genomic Mismatch Scanning (GMS) and Genome Mapping with DNA Microarrays

PARTICIPANT:

Basinsid, Margret, Lilly Research Laboratories, IN (MBB@Lilly.com)

Berghmans, Stephane, University of Liege, Belgium (stephane@stat.fmv.ulg.ac.be)

Champagne, Guy, DNA LandMarks, Inc., Canada (champagg@magellan.umontreal.ca)

Di Girolamo, Maria, Consorzio Mario Negri Sud, Italy

Ebeling, Saskia, Netherlands Red Cross, The Netherlands (ebeling@tip.nl)

Goff, Stephen, Novartis, Research Triangle Park, N.C. (goff@am.abru.cg.com)

Hornum, Lars, Hagedorn Research Institute, Denmark (laho@hrl.dk)

Jeon, Jin-Tae, Swedish Univ. of Agricultural Sci., Sweden (Jintae.Jeon@bmc.uu.se)

Lidmer, Ann-Sofie, Astra Hassle AB, Sweden (Ann-Sofie.Lidmer@hassie.se.astra.com)

Lodhi, Muhammad, Cold Spring Harbor Laboratory (lodhi@cshl.org)

Morshed, Syad Ahmed, Yale University (Morshed@kms.ac.sp.om) (Morshedsa@aol.com)

Parkin, Isobel, John Innes Center, United Kingdom (Isobel.Parkin@bbsrc.ac.uk)

Podlutsky, Andrej, KaŁolinska Institute, Sweden (andrew.podlutsky@cnt.ki.se)

Reddy, Ambati, University of Michigan (APReddy@umich.edu)

Rogel-Gaillard, Claire, Inst.National de la Recherche Agronomique, France (rogel@biotec.jouy. inra.fr)

Zhang, Y. Hi, University of Massachusetts (yhzhang@umbsky.cc.umb.edu)