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文献和实验ampicillin at 100 ug/ml) and incubate at 37deg C 8-10 hours with shaking at 250 rpm. Transfer the culture to an Ehrlenmeyer flask containing 50 ml of similar media, and incubate further for 11-14 hours. 2. Harvest the cells by centrifugation at 3000 rpm
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
Protocol for Reverse Transcription and Amino-allyl Coupling of RNA
primer, mix 1-2 mg mRNA with 5 ug of anchored oligo-dT [(dT)20-VN] (Operon, HPLC purified) in a total volume of 18 mL. One reaction for sample mRNA and one for reference mRNA. oligo dT 5 mg of 2.5 mg/mL 2 mL mRNA/water 1-2
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