Patent ID: 7700289
Filing Date: 2010-04-20
Classification: C12P,C12Q

Abstract:
1. A method of labeling RNA in a sample comprising: a) contacting a sample comprising a plurality of RNAs with calf intestinal alkaline phosphatase under conditions and for a time sufficient to result in dephosphorylated RNA; b) mixing the dephosphorylated RNA with DMSO to produce a resultant solution comprising DMSO at a concentration in the range from about 20% to about 70% and heating the resultant solution in a temperature range of about 65° C. to about 100° C., followed by cooling the resultant solution to about 0° C. to about 10° C.; c) contacting the resultant solution with an enzyme having RNA ligation activity in the presence of a labeled substrate under conditions sufficient to result in coupling of the labeled substrate to the dephosphorylated RNA to provide labeled RNA, said coupling occurring in the presence of an inactivated calf intestinal alkaline phosphatase no longer having RNA phosphatase activity; and wherein no phenol extraction is done after said contacting step a) and before said contacting step c); and wherein steps a), b), and c) all occur in the same reaction vessel.