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文献和实验Chromatin immunoprecipitation on native chromatin from cells and tissues
fraction P, in 50µl of dialysis buffer and store at 4°C. Quality Control of Chromatin Take the Optical Density (OD) of each fraction at 260nm; In separate 1.5mL Eppendorf tubes, put 0.5µg of each fraction (S1, S2, and P);
Revised amino-allyl labeling protocol
and the Howard Hughes Medical Institute http://cmgm.stanford.edu/pbrown/protocols/amino-allyl.htm A. RT Reaction 1. To anneal primer, mix 1-2 m g mRNA with 5 ug of anchored oligo-dT [(dT)20 -VN] (Operon, HPLC purified) in a total volume of 18 m L
(2)反应体系(25μL): F引物 0.5μL R引物 0.5μL SuperMix 12.5μL Passive Reference Dye 0.5μL cDNA 2μL ddH2O 9μL (3)程序设置如下(Eppendorf PCR仪): 95℃ 3min 95℃ 20s 55℃ 20s 40循环 72℃
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