Patent ID: 8067174
Filing Date: 2011-11-29
Classification: C12N

Abstract:
1. A polymerase chain reaction (PCR) method for amplifying a DNA template comprising, (a) providing a composition comprising an immobilized DNA polymerase or catalytically active fragment thereof each having an active site and an immobilization site, the immobilized DNA polymerase or the catalytically active fragment further comprising a linker, a matrix molecule, and a substrate material, wherein, i) the DNA polymerase or the catalytically active fragment is bonded by a covalent bond to the linker, ii) the linker is bonded by a chemical bond to the substrate material, iii) the immobilization site of the DNA polymerase or the catalytically active fragment is distant from the active site, and iv) the matrix molecule comprises a non-reactive terminal group and is connected to the substrate material together with the linker, and further wherein the average number of covalent bonds formed between the immobilized DNA polymerase or the catalytically active fragment and the linker is such that the average activity of the immobilized DNA polymerase or the catalytically active fragment is more than about 10% of the activity of solution phase DNA polymerase, (b) contacting the composition with a DNA template and amplification reagents; and (c) amplifying the DNA template with the immobilized DNA polymerase or the catalytically active fragment bound to the composition under PCR conditions to replicate the DNA template and further wherein the catalytically active fragment has at least about 70% of the activity of the DNA Polymerase.