Patent ID: 8822670
Filing Date: 2014-09-02
Classification: C12N,C12P,C12Q

Abstract:
1. A method of synthesizing a cDNA chain using an insoluble carrier having on the surface thereof a copolymer composed of a first monomer having a 2-methacryloyloxyethyl phosphorylcholine (MPC) group, a second monomer having p-nitrophenyloxycarbonyl polyethylene glycol methacrylate (NPMA) group and a third monomer having a butyl methacrylate (BMA) group, comprising: (a) allowing at least one polynucleotide for DNA elongation, containing a sequence complementary to the poly-A tail of an mRNA, to bind to said insoluble carrier, wherein the polynucleotide has an amino group; (b) adding a sample solution containing an mRNA having the poly-A tail, a solution containing a reverse transcriptase, and a solution containing nucleotide monomers, to said carrier having immobilized thereon the nucleotide for DNA elongation to thereby form a liquid phase containing said carrier; (c) allowing said poly-A tail of the mRNA to hybridize with the sequence complementary to the poly-A tail in said liquid phase obtained in step (b); and (d) allowing the polynucleotide for DNA elongation to elongate using the mRNA as a template, to thereby form an antisense cDNA complementary to the mRNA in said liquid phase obtained in step (b), wherein said polynucleotide for DNA elongation is immobilized onto the surface of the carrier via a covalent bond formed by the reaction of the amino group of the polynucleotide with the p-nitrophenyloxycarbonyl group of the NPMA contained in the copolymer with leaving of the p-nitrophenyloxy group of the p-nitrophenyloxycarbonyl group.