ISOTHERMAL AMPLIFICATION OF NUCLEIC ACIDS WITHIN A POROUS MATRIX

摘要

Provided herein are methods for amplification a target dsDNA that is impregnated within a porous matrix using endonuclease-assisted DNA amplification. The amplicons may be subsequent detected within the porous matrix or may be eluted out of the porous matrix. Methods for extracting a genetic material from a biological sample using endonuclease-assisted DNA amplification within a porous matrix are also provided.

主权项

1. A method of producing at least one amplicon based on a target double stranded DNA within a porous matrix comprising:
(a) providing the porous matrix; (b) impregnating the target double stranded DNA within the porous matrix; (c) contacting the impregnated, target double stranded DNA with a DNA amplification reaction mixture comprising at least one inosine-containing primer, at least one 5′→3′ exonuclease-deficient DNA polymerase having strand displacement activity, at least one nuclease that is capable of nicking a DNA at a residue 3′ to an inosine residue, and a dNTP mixture; (d) amplifying at least one portion of the impregnated target double stranded DNA within the porous matrix using the DNA amplification reaction mixture of step (c) to produce the at least one amplicon within the porous matrix; and (e) determining a rate of production of the at least one amplicon within the porous matrix.