Chimera-free, high copy number YAC libraries and efficient methods of analysis. Technical progress report
Evidence is accumulating to indicate that the mouse genome YAC library constructed in a rad52 host strain is quite low in chimeric YACs and in unstable YACs. We have optimized the transformation efficiency of two recombination-deficient strains which are isogenic with CGY2570. They are the rad52 strain CGY2872 and the rad52 radl strain CGY2897. The genotype of these gene-disrupted strains was verified by testing the uv and MMS sensitivity of the strains. These recombination-deficient strains grow about 10% slower than the isogenic wild-type strain and transform somewhat less efficiently. However, by optimizing the parameters described above, we have obtained reproducible transformation efficiencies which are only about 4-fold and 8-fold poorer than that of the isogenic wild-type strain. Using this procedure, we have obtained over 5,000 YACs of about 200 kb average size in the rad52 host and several thousand YACs of about 400--500 kb average size in the rad52 and rad52 radl hosts. Therefore, all of the procedures are now in place to build a total human genome library in a recombination deficient host.
- Research Organization:
- Collaborative Research, Inc., Bedford, MA (United States)
- Sponsoring Organization:
- USDOE, Washington, DC (United States)
- DOE Contract Number:
- FG02-92ER61399
- OSTI ID:
- 10130102
- Report Number(s):
- DOE/ER/61399-1; ON: DE93008551
- Resource Relation:
- Other Information: PBD: [1992]
- Country of Publication:
- United States
- Language:
- English
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