By Alan S. Waldman
Provides major peer-reviewed protocols to hold out recombinant investigations within the lab. Divided into 3 components for simple studying and reference.
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Additional info for Genetic Recombination, Reviews and Protocols
The first reports describing I-SceI cleavage of mammalian chromosomes were in 1994– 1995 (14,15), and as with HO in yeast, I-SceI has since been used in a wide variety of HR and DSB repair studies in mammalian cells. Here we review the systems for delivering I-SceI nuclease to mammalian cells and the strategies for detecting various outcomes of DSB repair, focusing primarily on HR. For additional information about the use of I-SceI in mammalian cells, see the review by Jasin (16). Analysis of Recombination With I-SceI Nuclease 37 Fig.
Do not take the first flowers on the first inflorescence for crossing. They might be infertile, but do not wait too long because flowers become smaller and the failure rate higher. 5. An important point in the determination of recombination frequencies is the analysis on the population level as well as on the level of the individual organism. Without DSB induction, on average one to two recombination events per seedling can be detected. For a statistical analysis 30–100 seedlings should be Intrachromosomal Homologous Recombination 33 analyzed.
Miller, C. , and Nickoloff, J. A. (2003) The mutagenic potential of a single DNA double-strand break is not influenced by transcription. DNA Repair 2, 1147–1156. 42. , Brennemann, M. , Chen, D. , and Nickoloff, J. A. (2002) DNA-dependent protein kinase suppresses double-strand break-induced and spontaneous homologous recombination. Proc. Natl. Acad. Sci. USA 99, 3758–3763. 43. Brenneman, M. , Wagener, B. , Miller, C. , and Nickoloff, J. A. (2002) XRCC3 controls the fidelity of homologous recombination: roles for XRCC3 in late stages of recombination.