The mouse Y chromosome: enrichment, sizing, and cloning by bivariate flow cytometry

Genomics
D E BergstromE M Simpson

Abstract

In this report, we demonstrate the utility of interleukin-2 (IL-2) stimulation of spleen cell cultures and bivariate flow cytometry in the analysis and purification of the C57BL/6J mouse Y Chromosome. We determined that the DNA content of the C57BL/6J Y Chromosome is approximately 94.7 Mb, making it similar in size to human Chromosome 16 and significantly larger than previous estimates. In addition, we describe the bulk isolation of mouse Y Chromosomes and demonstrate enrichment of the isolated material using a fluorescence in situ hybridization strategy. We detail the construction of two small insert Y Chromosome-specific libraries, ideal for sampling Y Chromosome sequences. From these libraries 1566 clones were analyzed. We provide a detailed characterization of 103 clones, generating nearly 50 kb of sequence. For 30 of these clones, we identify regions of homology to known Y chromosomal sequences, confirming the enrichment of the sorted DNA. From the remaining characterized clones, we describe the development of 15 male-specific PCR assays and 19 male-female PCR assays potentially originating from the pseudoautosomal region or other areas of X-Y or autosome-Y homology.

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Citations

May 14, 2011·Molecular Ecology Resources·Maja P GremingerPeter Wandeler
Jul 12, 2002·Genesis : the Journal of Genetics and Development·Elizabeth M SimpsonCharles P Lerner
Mar 30, 2001·Nature Genetics·P J WangD C Page
Jul 25, 2007·The Journal of Heredity·Shu-Jin LuoStephen J O'Brien
Jul 12, 2007·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Sanjay Kumar, Selvarangan Ponnazhagan

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