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 |
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Academic Press Inc.
2014
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| 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 |
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| author | Osahor, Andrew N. Tan, Chau-Yan Sim, Edmund U. H. Lee, Choon-Weng Narayanan, Kulathuramaiyer |
| author_facet | Osahor, Andrew N. Tan, Chau-Yan Sim, Edmund U. H. Lee, Choon-Weng Narayanan, Kulathuramaiyer |
| author_sort | Osahor, Andrew N. |
| building | UNIMAS Institutional Repository |
| collection | Online Access |
| description | 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. |
| first_indexed | 2025-11-15T06:54:09Z |
| format | Article |
| id | unimas-17707 |
| institution | Universiti Malaysia Sarawak |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-15T06:54:09Z |
| publishDate | 2014 |
| publisher | Academic Press Inc. |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | unimas-177072017-09-21T02:34:32Z http://ir.unimas.my/id/eprint/17707/ Short homologies efficiently generate detectable homologous recombination events Osahor, Andrew N. Tan, Chau-Yan Sim, Edmund U. H. Lee, Choon-Weng Narayanan, Kulathuramaiyer QD Chemistry 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. Academic Press Inc. 2014 Article PeerReviewed text en http://ir.unimas.my/id/eprint/17707/1/Short%20homologies%20efficiently%20generate%20detectable%20homologous%20%28abstract%29.pdf Osahor, Andrew N. and Tan, Chau-Yan and Sim, Edmund U. H. and Lee, Choon-Weng and Narayanan, Kulathuramaiyer (2014) Short homologies efficiently generate detectable homologous recombination events. Analytical Biochemistry, 462. pp. 26-28. ISSN 0003-2697 http://www.sciencedirect.com/science/article/pii/S0003269714002498 doi : 10.1016/j.ab.2014.05.030 |
| spellingShingle | QD Chemistry Osahor, Andrew N. Tan, Chau-Yan Sim, Edmund U. H. Lee, Choon-Weng Narayanan, Kulathuramaiyer Short homologies efficiently generate detectable homologous recombination events |
| title | Short homologies efficiently generate detectable homologous
recombination events |
| title_full | Short homologies efficiently generate detectable homologous
recombination events |
| title_fullStr | Short homologies efficiently generate detectable homologous
recombination events |
| title_full_unstemmed | Short homologies efficiently generate detectable homologous
recombination events |
| title_short | Short homologies efficiently generate detectable homologous
recombination events |
| title_sort | short homologies efficiently generate detectable homologous
recombination events |
| topic | QD Chemistry |
| url | http://ir.unimas.my/id/eprint/17707/ http://ir.unimas.my/id/eprint/17707/ 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 |