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文献和实验The ribonuclease protection assay (RPA)
The ribonuclease protection assay (RPA) is a highly sensitive and specific method for the detection of mRNA species. The assay was made possible by the discovery and characterization of DNA-dependant RNA polymerases from the bacteriophages SP6, T
Cool growth competent E . coli
, while swirling, to final 7 % (~1.4 ml). -Place on ice, 10 min. Dispense into freezing tubes (~0.5-1 ml ea) -Freeze in liquid N2; transfer to -80°. 1 liter SOB: 20 g bactotryptone, 5 g bacto-yeast extract, 0.5g NaCl Dissolve all in 950 ml ddH
and to the vacuum. Filter the lysate through the Whatman,changing it often as it gets clogged. Filtrate may now be stored a 4℃ until it is time to run over an affinity column. Note: Don't store for more than a couple of days at 4℃. References K.D.Puri,E.B
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