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文献和实验Clonality - X Chromosome Inactivation Assay
(Life Technologies) 10X PCR buffer (100 mM Tris-HCL pH 8.3, 500 mM KCL, 15 mM MgCl2, 0,01 % w/v gelatin, autoclaved) (Perkin Elmer) dNTP, each 10 mM (Perkin Elmer) 7-deaza dGTP, 10 mM (Boehringer Mannheim) DMSO, minimum 99.5%, GC (Sigma
Single tube confirmation PCR protocol
Polymerase. - Add 35 µl of the PCR mix to each of the 20 PCR tubes that already contain the 15 µl of primers and template. 110 µl 5 µl 10 x Taq buffer(see below) 11 µl 0.5 µl 20 mM dNTP's (0.2 mM) 11 µl 0.5 µ
ES CELL DNA EXTRACTION: TUBE ME
off supernatant as in step 16. 19. Store the open tube on the bench at TRm until the last traces of fluid have evaporated. (i.e.: Air dry). 20. Dissolve the DNA in 30µl (or 50 µl, and use half of sample) of 10mM Tris pH 8.5 (or TE). (pipet
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