Course Announcement

[Karen Otto in the Meetings Office]

                     COLD SPRING HARBOR LABORATORY

                       SPRING COURSE ANNOUNCEMENT


PROTEIN PURIFICATION AND CHARACTERIZATION
April 8 - 22, 1991

Daniel R. Marshak, Cold Spring Harbor Laboratory
Bruce W. Erickson, University of North Carolina
Jim Kadonaga, University of California, San Diego
John A. Smith, Harvard Medical School

This course is intended for scientists who are not familiar with
techniques of protein isolation and characterization.  Students will
learn the major techniques in protein purification by actually
performing four separate isolations including: (i) a regulatory
protein from muscle tissue; (ii) a fusion protein from E. coli;
(iii) a DNA binding protein from nuclei of tissue culture cells; and
(iv) a chemically synthesized peptide.  A variety of
chromatographic, electrophoretic, and bulk fractionation techniques
will be employed including: ion exchange, gel filtration,
hydrophobic interaction, affinity-based adsorption, and
immunoaffinity chromatography; polyacrylamide gel and two-
dimensional gel electrophoresis and electroblotting; precipitation
by salt and pH; and HPLC analysis.  Methods of protein
characterization will be discussed including amino acid analysis,
protein sequencing, and mass spectrometry.  Emphasis will be placed
on methods of protein purification rather than automated
instrumental analysis.  Guest lecturers will discuss protein
structure, modifications of proteins, and methodologies for protein
purification.  Applications of protein biochemistry to various areas
of research in molecular biology will be discussed.  Guest lecturers
include: R. Aebersold, L. Gierasch, G. Hart, Y. Paterson, N. Pace,
G. Rose, and K. Wilson.

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CLONING & ANALYSIS OF LARGE DNA MOLECULES
April 8 - 22, 1991

Bruce Birren, California Institute of Technology
Sue Klapholz, Cell Genesys, Inc.
Nancy Shepherd, E. I. du Pont de Nemours & Co.

This course will cover the theory and practice of manipulating and
cloning high molecular weight DNA.  Lectures and laboratory work
will deal with the use of bacteriophage P1 and yeast artificial
chromosome (YAC) cloning systems, the isolation and manipulation of
high molecular weight DNA from mammalian cells for cloning
(including the size-selection of >200 kb DNA fragments), and the
analysis of high molecular weight DNA by pulse field gel (PFG)
separation techniques.  P1 and YAC recombinant DNA molecules will be
produced, introduced into cells (E. coli and yeast, respectively),
and reisolated after appropriate clone selection and colony
screening procedures.  A variety of size standards for pulsed-field
gel electrophoresis will be prepared and gels will be run to compare
the DNA separation capabilities of the common PFG techniques.
Students will gain experience with physical mapping of YAC inserts
and high molecular weight genomic DNA.  Lectures by outside speakers
on topics of current interest will supplement the laboratory work.

Application deadline:  January 15, 1991

Tuition, Room and Board  . . . . . . . . . . . . . . . . . . $1,475

Application/information may be obtained from:

REGISTRAR
Cold Spring Harbor Laboratory
Bungtown Road
Cold Spring Harbor, NY  11724

EMAIL:  needinfo at cshl.org
        (include your name & address as well as course name)
PHONE:  (516) 367-8346
FAX:    (516) 367-8845