Patent ID: 7176025
Filing Date: 2007-02-13
Classification: C12N,C12Q

Abstract:
1. A method for generating a hybrid nucleic acid, said method comprising: (a) preparing a complex of first and second primer extension products said complex comprising a 3′ single stranded portion, wherein said complex comprising a 3′ single stranded portion is prepared according to a method comprising: (i) extending a first primer hybridized to a target RNA with an enzyme comprising RNA-dependent DNA polymerase activity, wherein the first primer is a composite primer comprising an RNA portion and a 3′ DNA portion, whereby a complex comprising a first primer extension product and the target RNA is produced; (ii) cleaving target RNA in the complex of step (i); (iii) extending a second primer hybridized to the first primer extension product with an enzyme comprising DNA-dependent DNA polymerase activity and an enzyme comprising RNA-dependent DNA polymerase activity, whereby a second primer extension product is produced to form a complex of first and second primer extension products; (iv) cleaving RNA from the first primer in the complex of step (iii) with an enzyme that cleaves RNA from an RNA/DNA hybrid such that a complex comprising hybridized first and second primer extension products comprising a 3′ single stranded portion is generated; and (b) hybridizing the 3′ single stranded portion of the complex of first and second primer extension products with a recipient nucleic acid molecule; whereby a hybrid nucleic acid is generated.