Chapter title |
A Method to Capture Large DNA Fragments from Genomic DNA
|
---|---|
Chapter number | 38 |
Book title |
Pseudomonas Methods and Protocols
|
Published in |
Methods in molecular biology, January 2014
|
DOI | 10.1007/978-1-4939-0473-0_38 |
Pubmed ID | |
Book ISBNs |
978-1-4939-0472-3, 978-1-4939-0473-0
|
Authors |
Geneviève Ball, Alain Filloux, Romé Voulhoux, Ball, Geneviève, Filloux, Alain, Voulhoux, Romé |
Abstract |
The gene capture technique is a powerful tool that allows the cloning of large DNA regions (up to 80 kb), such as entire genomic islands, without using restriction enzymes or DNA amplification. This technique takes advantage of the high recombinant capacity of the yeast. A "capture" vector containing both ends of the target DNA region must first be constructed. The target region is then captured by co-transformation and recombination in yeast between the "capture" vector and appropriate genomic DNA. The selected recombinant plasmid can be verified by sequencing and transferred in the bacteria for multiple applications. This chapter describes a protocol specifically adapted for Pseudomonas aeruginosa genomic DNA capture. |
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