File:Homologous Recombination Deletion method one.png: Difference between revisions
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==Image Explanation== | |||
Linear sequences of DNA were created using PCR from synthetic primers (the red stretch of DNA). Regions A, B and C inside the target region inside the genome (green) and inside the linear PCR sequence (red) are homologous regions. Cm^R is antibiotic resistance, S indicates an I-ScsI cleavage site. | Linear sequences of DNA were created using PCR from synthetic primers (the red stretch of DNA). Regions A, B and C inside the target region inside the genome (green) and inside the linear PCR sequence (red) are homologous regions. Cm^R is antibiotic resistance, S indicates an I-ScsI cleavage site. | ||
Revision as of 19:35, 10 February 2013
Image Explanation
Linear sequences of DNA were created using PCR from synthetic primers (the red stretch of DNA). Regions A, B and C inside the target region inside the genome (green) and inside the linear PCR sequence (red) are homologous regions. Cm^R is antibiotic resistance, S indicates an I-ScsI cleavage site.
The DNA is electroporated into the cell, then a plasmid coding for recombinase elements used for recombination. Cells that succeed in recombination are isolated.
Another plasmid coding for I-SceI meganuclease was inserted into the cell to remove the Cm^R gene. Then, a version of Rec-A mediated DSB recombinational repair was used to ligate the ends together.
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| current | 19:33, 10 February 2013 |  | 1,184 × 1,829 (229 KB) | Evan J. Weaver (talk | contribs) |
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