Application of Interspersed Repetitive Sequence Polymerase Chain Reaction for Construction of Yeast Artificial Chromosome Contigs

Methods ◽  
1997 ◽  
Vol 13 (4) ◽  
pp. 327-335 ◽  
Author(s):  
Kent Hunter
Keyword(s):  
Polymerase Chain Reaction ◽  
Repetitive Sequence ◽  
Yeast Artificial Chromosome ◽  
Artificial Chromosome ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals Alu polymerase chain reaction: a method for rapid isolation of human-specific sequences from complex DNA sources

1989 ◽  
Vol 86 (17) ◽  
pp. 6686-6690 ◽  
Author(s):  
D L Nelson ◽  
S A Ledbetter ◽  
L Corbo ◽  
M F Victoria ◽  
R Ramírez-Solis ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Human Genome ◽  
Yeast Artificial Chromosome ◽  
Hybrid Cell ◽  
Artificial Chromosome ◽  
Chain Reaction ◽  
General Applicability ◽  
Simple Method ◽  
Alu Repeat ◽  
Polymerase Chain

Current efforts to map the human genome are focused on individual chromosomes or smaller regions and frequently rely on the use of somatic cell hybrids. We report the application of the polymerase chain reaction to direct amplification of human DNA from hybrid cells containing regions of the human genome in rodent cell backgrounds using primers directed to the human Alu repeat element. We demonstrate Alu-directed amplification of a fragment of the human HPRT gene from both hybrid cell and cloned DNA and identify through sequence analysis the Alu repeats involved in this amplification. We also demonstrate the application of this technique to identify the chromosomal locations of large fragments of the human X chromosome cloned in a yeast artificial chromosome and the general applicability of the method to the preparation of DNA probes from cloned human sequences. The technique allows rapid gene mapping and provides a simple method for the isolation and analysis of specific chromosomal regions.

Sign in / Sign up

Export Citation Format

Share Document