Deriving probes from large-insert clones by PCR methods.
This unit describes several polymerase chain reaction (PCR)-based methods to obtain DNA fragments from clones with large inserts without prior knowledge of the insert DNA sequence. The protocols can be categorized into three groups: (1) methods to generate DNA fragments at random representing the entire length of the cloned insert, (2) methods to generate DNA fragments representing the extremities of an insert, and (3) methods to generate complex probes suitable for fluorescence in situ hybridization. Support protocols describe direct cloning of these PCR products and the isolation of total yeast DNA from yeast artificial chromosome (YAC) clones.
Albertsen
H
H
University of Utah, Salt Lake City, Utah, USA.
Thliveris
A
A
Riley
J H
JH
Munroe
D J
DJ
Watkins
P
P
Basson
C T
CT
D020087:Alu Elements; D027042:Chromosomes, Artificial, P1 Bacteriophage / Q000235:genetics; D018244:Chromosomes, Artificial, Yeast / Q000235:genetics; D003001:Cloning, Molecular; D015342:DNA Probes / Q000235:genetics / Q000302:isolation & purification; D005822:Genetic Vectors; D005826:Genetics, Medical; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D015336:Molecular Probe Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*
D015342:DNA Probes