Patent ID: 7824890
Filing Date: 2010-11-02
Classification: C12Q

Abstract:
1. An isothermal process for amplifying a single stranded nucleic acid template which comprises: (a) applying a forward primer (primer-1) to the 3′ region of the template; (b) extending said primer by a polymerase; (c) removing or degrading the 5′ terminus of said primer to leave a partly degraded duplex product (product-1); (d) applying a further primer-1 molecule (primer-1a) to the region exposed by degradation of the 5′ terminus of the extended primer (primer-1) so that the 5′ region of said further primer (primer-1a) binds to the template; (e) allowing strand invasion of product-1 by the 3′ terminus of said further primer (primer-1a), whereby the 3′ terminus of said further primer hybridises with the template in place of the 5′ terminus of product-1; (f) extending said primer-1a by a strand displacing polymerase causing release of product-1 (amplicon) from the template; and (g) reacting product-1 with a reverse primer (primer-2) which binds to the 3′ region of product-1; (h) repeating steps (b) to (g) where the forward primer (primer-1) is replaced by the reverse primer (primer-2) in order to produce product-2 (amplicon); wherein the original template is re-created by the reaction between product-2 and the 3′ terminus of primer-1, whereby the 5′ terminus of the primer is extended onto by the 3′ terminus of the template.