Patent ID: 6238865
Filing Date: 2001-05-29
Classification: C12P

Abstract:
A method for modifying a 3'-terninus of a preselected RNA sequence so as to produce a 3'-terminus modified preselected RNA sequence having at least a single modified nucleotide mono-phosphate at said 3'-terminus, comprising the steps of:a. preparing a reaction mixture which comprises a modified nucleotide tri-phosphate selected from the group consisting of dATP, dGTP, dCTP, TTP, ATP, GTP, CTP, UTP and analogs thereof, a nucleotide template having a sequence complementary to the 3'-terminus portion of said preselected RNA sequence and a 5'-overhang sequence, said 5'-overhang sequence including at least a first overhang nucleotide mono-phosphate complementary to said modified nucleotide tri-phosphate, an enzyme capable of elongating said 3'-terminus of said preselected RNA sequence in a template dependent fashion to produce a sequence complementary to said nucleotide template, and said preselected RNA sequence to be modified at said 3'-terminus; andb. incubating said reaction mixture at 16-45.degree. C. for at least 2 minutes to produce said 3'-terminus modified preselected RNA sequence.