course in YAC analysis

Geordie +1865 740 011 ayoung at vax.oxford.ac.uk
Mon Mar 6 07:21:36 EST 1995


        WELLCOME SUMMER SCHOOLS : FIFTHTEENTH ADVANCED COURSE

              HUMAN GENOME ANALYSIS : FROM YAC TO GENE

                     27th July - 4th August 1995

Division of Biochemistry & Molecular Biology, United Medical and Dental 
Schools of Guy's and St Thomas's Hospitals, Guy's Campus, London

All email enquiries to: wss at umds.ac.uk

An intensive laboratory course to introduce participants to YAC and cosmid 
contig assembly, clone characterisation and gene hunting.

Programme

1.	Analysis of YAC clones 

A range of methods for analysis of individual YAC clones will be covered.  
These include the preparation of YAC plugs, gel purification of artificial 
chromosomes, experiments to recover sequences from within and at the ends of 
YAC clones for further studies.

2.	Subcloning of YACs and integration with prokaryotic cloning system

For more detailed mapping and isolation of specific landmarks such as genes 
and polymorphisms it is necessary to integrate YACs with prokaryotic cloning 
systems containing smaller inserts. Direct screening of cosmid arrays with 
YACs, and subcloning of individual YACs into lambda phage and cosmid vectors 
will be carried out as part of the course.

3.	Assembly of chromosome specific cosmid contigs	

Cosmid library screening techniques will be discussed.  Filters containing 
hundreds of cosmids will be screened with YACs.  YAC and cosmid based 
techniques will be used to generate cosmid contigs.

4.	FISH analysis of YAC and cosmid clones

YACs and cosmids will be mapped on metaphase chromosomes.  Selected cosmid 
clones will be mapped on extended chromatin fibers for high resolution.

5.	Retrofitting and fragmentation of YACs

Manipulation of YAC inserts e.g. the joining of two or more inserts by 
recombination between YACs, or the introduction of individual YACs into 
heterologous cells for further study of the function of the cloned 
sequences, may require alteration of the selectable markers in the YAC. 
Methods will be used to retrofit such markers into the vector sequences. 
Recombination in yeast can also be used to fragment YACs and generate a 
nested set of shorter artificial chromosomes, as well as to amplify the YAC 
DNA over the yeast chromosomal background.  YAC fragmentation and 
amplification experiments will be performed.  All these methods will also 
provide experience in yeast transformation.

6.	Application of YACs to specific problems in human genetics

The particular features of YAC cloning have resulted in the development of 
new approaches to solving problems in human genetics. Specific examples will 
be presented both as part of the experimental work and in discussion.

7.	Gene hunting

Experiments will include cDNA and non cDNA based approaches.  cDNA 
libraries will be screened with cosmid genomic DNA inserts.  In addition 
cDNAs will be isolated using cDNA selection techniques.  These techniques 
will be compared with exon trapping, a technique which does not require 
prior knowledge of the expression pattern of a gene.

8.	YAC transfer to mammalian cells

YAC transfer to mammalian cells by spheroplast fusion will be undertaken 
using a short term assay to determine transfer efficiency.

Informal tutorials and evening lectures (by invited speakers) will 
supplement the laboratory sessions.

Course Instructors

DUNCAN CAMPBELL (Oxford), ANGELA DAVIES (London),  DAVID MARKIE (London), 
TONY MONACO (Oxford), IOANNIS RAGOUSSIS (London), CAROL WISE (Texas).
	
Confirmed speakers include:

ANDREA BALLABIO (Milan) ADRIAN BIRD (Edinburgh), ALAN BUCKLER 
(Massachusetts),  IAN DUNHAM (Hinxton), MICHAEL LOVETT  (Texas), CRAIG 
VENTER (Maryland).

Participants

Applicants should be post-doctoral (or close to) scientists engaged in 
relevant research.  Applicants with molecular biology and/or some cell 
culture experience will be particularly welcome.  The course is subsidised 
by the Wellcome Trust for scientists based in academic institutions anywhere 
in the world.  This is a residential course, without exception, and there is 
a charge of 400 pounds towards board and lodging.

Applications

There are no formal application forms, but applicants should send a copy of 
their full CV together with a 300 word outline of their research plans to Dr 
Pelin Faik, Course Co-ordinator, Division of Biochemistry & Molecular 
Biology, UMDS, Guy's Campus, London Bridge SE1 9RT.

Tel:  0171 403 6998 	Fax:   0171 407 5281   Email: wss at umds.ac.uk

	Closing date for applications is 31st March 1995.
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    /   \   /                 |    /      Alan Young @ uk.ac.ox.vax
   /____/  /  ___    __       |___/
  /    /  /  /   |  /  |         /           "The bigger they are,
 /     \_/\__\__/|_/  /_________/               the harder they fall on you."



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