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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
50 µl total volume *10 x Taq buffer contains: 100 mM Tris-HCl (pH 8.4), 500 mM KCl, 15 mM MgCl2. *The final concentrations are shown in parentheses. *The Taq Polymerase should be added last and PCR mixture should be kept on ice
Purification of Plasmid from 50 ml-culture
of cold 70% ethanol. Discard the fluid. 14. Add to the pellet 400 ul of TE containing DNase-free RNase A (20 ug/ml). Incubate the tube for 30 min at 37 C. 15. After 30 min, carefully check the content of the tube. If nucleic acid pellet
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