Chromosome, Microdissection and Microcloning
One million base pairs of nucleotides.
Laser-capture microdissection. A devise that delivers laser pulses to dissect tissues or
chromosomes. Pulses can be tuned in variable micron diameter.
Metaphase Chromosomes
Condensed chromosomes at metaphase during the cell cycle.
A technology that assesses gene expression by arraying PCR-ampliFed cDNA clones or
genes at high density on derivatized glass microscope slides (known as a DNA chip).
Relative expression levels are simultaneously determined by speciFc probe
A method to physically cut and collect chromosomal fragments by means of very Fne
needles and micromanipulators or by a laser microbeam.
Polymerase Chain Reaction(PCR)
A technique by which target DNA sequences may be replicated (ampliFed) selectively
in vitro
The global analysis of expressed proteins in a cell or tissue phenotype in attempt to
establish the relationship between the genome sequence, the expressed proteins, and
the proteinā€“protein interactions.
A yeast artiFcial chromosome based cloning vector capable of accommodating larger
pieces of genomic DNA (average insert
100 kb).
This article reviews the pertinent aspects of chromosome structure, microdissection
techniques, and methods of microcloning. Recent applications of this technology
are discussed. Microdissection is a specialized aspect of cell microsurgery by
which one can remove a chromosome fragment from a cell in metaphase. The DNA
material thus obtained can be used for molecular cloning experiments. Coupling this
technology with
in vitro
DNA ampliFcation using polymerase chain reaction (PCR)
and gene-transfer techniques have made possible the performance of biochemical
and biological experiments using small amounts of DNA obtained from chromosome
fragments. The emerging technology of laser-capture microdissection (LCM) and
its potential applications in global gene expression and proFling (proteomics) is
also introduced.
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