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文献和实验RNA Purification Protocol for 10-20mg of Animal Tissue
# SSI-1015-39). 3. Add 200 μl of PNP Buffer to the cell lysate. Mix the sample gently by inverting the tube 10 times. 4. Place the tube on ice for 5 minutes. 5. Centrifuge at max speed (.13,000 x g) for 3 minutes at room temperature
Reverse Transcription of RNA- 20l rxn
for 10mins. Notes All reagents should be diluted in dH20 that has been dep''d and autoclaved. Add all reagents except the RT and RNA and let them sit in the presence of the RNase for 15 mins. I use 1.5 ug of Poly A+ RNA in this rxn
TOP10 chemically competent cells
10 mM KOAc pH 7.0 (10 ml of a 1M stock/L) 80 mM CaCl2 .2H2 O (11.8 g/L) 20 mM MnCl2 .4H2 O (4.0 g/L) 10 mM MgCl2 .6H2 O (2.0 g/L) 10% glycerol (100 ml/L) adjust pH DOWN to 6.4 with 0.1N HCl if necessary
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