By Andrew Ward
Univ. of tub, united kingdom. Brings jointly the basic molecular, genetic, and embryological tools utilized in latest laboratories for the identity and research of imprinted genes. every one method is defined intimately. define layout.
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And Surani, M. A. (1995) Peg1/Mest imprinted gene on chromosome 6 identified by cDNA subtraction hybridization. Nat. Genet. 11, 52–59. 17. , Barton, S. , Surani, M. , and Ishino, F. (1998) Identification of the Meg1/Grb10 imprinted gene on mouse proximal chromosome 11 a candidate for the Silver-Russell syndrome gene. Proc. Natl. Acad. Sci. USA 95, 1102–1107. 18. Kelsey, G. and Reik, W. (1998) Analysis and identification of imprinted genes. Methods 14, 211–234. 19. , Wroe, S. , Wells, C. , Miller, H.
Searle, A. G. and Beechey, C. V. (1990) Genome imprinting phenomena on mouse chromosome 7. Genet. , Camb. 56, 237–244. 37. McLaughlin, K. , Szabó, P. , and Mann, J. R. (1996) Mouse embryos with paternal duplication of an imprinted chromosome 7 region die at midgestation and lack placental spongiotrophoblast. Development 122, 265–270. 38. Epstein, C. J. (1986) Developmental genetics. Experientia 42, 1117–1128. 39. , Schnieke, A. , Scott, A. , and Kind, A. J. (1996) Selective ablation of differentiated cells permits isolation of embryonic stem cell lines from murine embryos with a non-permissive genetic background.
Slit uterus open and remove up to 10 fetuses. Place them into a fresh 10-cm plate with 10 mL DPBS+. Using #5 watch maker’s forceps, pinch off the head and remove liver from each fetus. 4. Transfer the carcasses to a fresh 10-cm plate containing 10 mL DPBS+. Tilt the plate, then, with a 1-mL syringe fitted with an 18-gage needle, draw up each fetus individually into the syringe to shear it into small pieces, then expel into a fresh 10-cm plate. 5. Aspirate DPBS+ from plate, add 10 mL trypsin-EDTA, then leave plate in incubator for 15 min.