Short homologies efficiently generate detectable homologous recombination events
When recombineering bacterial artificial chromosomes (BACs), it is common practice to design the ends of the donor molecule with 50 bp of homology specifying its insertion site. We demonstrate that desired recombinants can be produced using intermolecular homologies as short as 15 bp. Although the...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Academic Press Inc.
2014
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| Subjects: | |
| Online Access: | http://ir.unimas.my/id/eprint/17707/ http://ir.unimas.my/id/eprint/17707/1/Short%20homologies%20efficiently%20generate%20detectable%20homologous%20%28abstract%29.pdf |
| Summary: | When recombineering bacterial artificial chromosomes (BACs), it is common practice to design the ends
of the donor molecule with 50 bp of homology specifying its insertion site. We demonstrate that desired
recombinants can be produced using intermolecular homologies as short as 15 bp. Although the use of
shorter donor end regions decreases total recombinants by several fold, the frequency of recombinants
with correctly inserted donor molecules was high enough for easy detection by simple polymerase chain
reaction (PCR) screening. This observation may have important implications for the design of oligonucleotides
for recombineering, including significant cost savings, especially for high-throughput projects that
use large quantities of primers. |
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