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文献和实验E.Z.N.A. Cycle-Pure Kit Spin Protocol
of the sample at 260nm and then at 280nm. The DNA concentration is calculated as follows: 260 DNA concentration = A x 50 x (Dilution Factor) ug/ml Fragments greater than 500bp in length can routinely be purified at > 80% yield. 260 Bands ranging
You will need for 1° cDNA synthesis: 5 x SuperScript RTase buffer (Gibco-BRL) 5mM methyl dNTPs (Pharmacia, substitute 5m dCTP for dCTP) 100mM DTT (Gibco-BRL) oligo dT primer/adaptor [a-32P] dATP (~400Ci/mmol, Amersham or DuPont) Ribonuclease inhibitor
you monitor every step using a phase contrast microscope (see below). 1 hour before nucleolar isolation, replace with fresh, pre-warmed medium. 2. Harvest cells by trpysinization. Rinse each dish 3X with pre-warmed PBS, and on removal of the last rinse
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