PRIMER SINGLE-STRANDED DNA, METHOD FOR PREPARING DOUBLE- STRANDED CDNA USING THE SAME, AND METHOD FOR AMPLIFYING ONE-SIDED SINGLE-STRANDED DNA

摘要

PROBLEM TO BE SOLVED: To synthesize each cDNA from various mRNAs in a trace amount of a specimen preserving an information concerning the directionality of a gene, to evenly amplify these, and to further amplify only one-sided single strand of the amplified double-stranded cDNA. ;SOLUTION: This method for preparing a one-sided single-stranded DNA of double-stranded DNA comprises the steps of: (a) hybridizing a primer single- stranded DNA designed to inhibit an exonuclease activity of a DNA polymerase to a poly A end of a mRNA, (b) linking an adaptor double-stranded DNA to one side of the double-stranded DNA, (c) amplifying the double-stranded cDNA by the elongation using two single-stranded DNAs constituting the adaptor double-stranded DNA as primers, and (d) asymmetrically amplifying a one-sided single-stranded DNA of the amplified double-stranded cDNA by the PCR using a primer single-stranded DNA hybridizing to a dA stretch of a double-stranded DNA and the adaptor double-stranded DNA.;COPYRIGHT: (C)2001,JPO