caa a22 4500 469062584 CHVBK 20180323132901.0 cr unu---uuuuu 170328e19920201xx s 000 0 eng 10.1007/BF00431250 doi (NATIONALLICENCE)springer-10.1007/BF00431250 Genomic mapping within the albino-deletion complex using individual early postimplantation mouse embryos [Elektronische Daten] [Shyam Sharan, Bernadette Holdener-Kenny, David Threadgill, Terry Magnuson] Sensitive methods for analysis of DNA from limited amounts of tissue are often difficult, error prone, and time consuming. Here, we describe a procedure for molecular analysis of individual early post-implantation mouse embryos by Southern analysis. The procedure involves embedding single embryos in agarose before lysing and deproteinizing in situ. The embedded DNA can be digested with restriction enzymes and analyzed by standard Southern-blotting procedures. The procedure is sensitive enough to detect single-copy sequences in embryos as early as day 6.5 of development. We have used the technique to genotype embryos homozygous for an embryonic lethal deletion. Normally, the lethal phenotype associated with such mutations is identified by a retrospective statistical analysis of abnormal embryos produced from a heterozygous cross as compared to those produced from a control cross. Now, if associated with a detectable DNA abnormality, the mutant embryo can be genotyped directly. We also report the use of this method for mapping cloned markers relative to deletion breakpoints. This approach can save considerable time since mapping would conventionally be done using restriction fragment length polymorphisms (RFLPs) detected in Mus musculus/Mus spretus interspecies hybrids. Using this procedure, we have been able to redefine the distal limits of the region of Chromosome (Chr) 7 containing a gene (eed) needed for development of the embryonic ectoderm. Springer-Verlag New York Inc, 1992 Sharan Shyam Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut Holdener-Kenny Bernadette Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut Threadgill David Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut Magnuson Terry Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut Mammalian Genome Springer-Verlag 3/2(1992-02-01), 79-83 0938-8990 3:2<79 1992 3 335 https://doi.org/10.1007/BF00431250 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00431250 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Sharan Shyam Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut NATIONALLICENCE 700 1- Holdener-Kenny Bernadette Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut NATIONALLICENCE 700 1- Threadgill David Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut NATIONALLICENCE 700 1- Magnuson Terry Department of Genetics, Case Western Reserve University, 2109 Adelbert Road, 44106, Cleveland, Ohio, USA aut NATIONALLICENCE 773 0- Mammalian Genome Springer-Verlag 3/2(1992-02-01), 79-83 0938-8990 3:2<79 1992 3 335 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer