17
Chromosome, Microdissection
and Microcloning
Nabil G. Hagag
StateUn
ivers
ityofNewYork,StonyBrook,NY,USA
1
Chromosome Structure and Organization
20
2
Preparation of Chromosomes for Microdissection
22
2.1
Aspects of Preparing Mammalian Chromosomes
22
2.1.1
Sources of DNA
22
2.1.2
Hypotonic Solution
23
2.1.3
Chromosome Fixation
23
2.1.4
Aging
24
2.1.5
GTG Banding for Chromosome Microdissection
24
2.2
Preparation of Dipteran Chromosomes
24
3
Methods of Chromosome Microdissection
25
3.1
Equipment
25
3.2
Methods
26
3.2.1
Oil Chamber Method
26
3.2.2
Videomicroscope Method
27
3.2.3
Laser Microbeam Method
29
3.3
Dissection and Collection of Chromosome Fragments by
the Videomicroscopy Method
30
4
Methods of Cloning and Analysis of Microdissected DNA
31
4.1
Cloning Methods
31
4.1.1
Microcloning
33
4.1.2
Linker Adapter with PCR Ampli±cation
34
4.1.3
PCR Ampli±cation of Chromosomal DNA
35
4.2
Analysis of Cloned DNA
38
5
Laser-capture Microdissection (LCM)
40
Encyclopedia of Molecular Cell Biology and Molecular Medicine, 2nd Edition
. Edited by Robert A. Meyers.
Copyright
2004 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.
ISBN: 3-527-30545-9
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