DIRECTED DNA CO-POLYMER MASS AMPLIFICATION FOR ssDNA DETECTION

摘要

The disclosure generally relates to the use of a bio-barcode (BBC) method for capture and demonstrates hybridization co-polymerization amplification readout of the BBC as a method of rapid detection of single stranded DNA output with an increased sensitivity for the initial target input to the BBC assay. The BBC assay reporter ssDNA can be modified for rapid readout. Two ssDNA molecules are designed to co-polymerize and then continually hybridize into double stranded DNA. The double stranded DNA can be optically and/or electrically detected. Kits related to the two ssDNA molecules and the BBC assay also are disclosed.

主权项

1. A method for making a double-stranded DNA molecule, the method comprising:
(a) providing a population of a first single-stranded DNA (ssDNA) oligonucleotide having a first region and a second region in the 5′ to 3′ direction, wherein the first region is 5′ in position relative to the second region; (b) providing a population of a second single-stranded DNA (ssDNA) oligonucleotide having a third region and a fourth region in the 3′ to 5′ direction for hybridization with the first ssDNA oligonucleotide, wherein:
(i) the third region is 3′ in position relative to the fourth region;(ii) the second region is complementary to the third region, and the first region is complementary to the fourth region; and(iii) the first region and the fourth region become overhangs when the second region and the third region hybridize together, and the second region and the third region become overhangs when the first region and the fourth region hybridize together; and (c) hybridizing the population of the first ssDNA oligonucleotide and the population of the second ssDNA oligonucleotide under a predetermined hybridization condition to form a double-stranded DNA (dsDNA) molecule, the dsDNA molecule having repetitive first ssDNA oligonucleotide units on one strand and repetitive second ssDNA oligonucleotide units on the other strand.