A process is disclosed for generating at least one partially double-stranded target nucleic acid, which contains at least one single-stranded region at a terminal end. The process comprises the steps of (a) providing at least one primer, P
1
, containing at least one labile nucleotide; (b) combining at least one target nucleic acid sequence with P
1
to generate a double-stranded polynucleotide containing at least one labile nucleotide; (c) exposing the double-stranded polynucleotide to conditions that promote single-strand cleavage of the polynucleotide at the site of the at least one labile nucleotide of primer P
1
; and (d) exposing the cleaved polynucleotide to conditions that promote the dissociation of the cleaved portions of primer P
1
from a terminal end. The labile nucleotide may be dUTP, wherein the single-stranded cleavage of the polynucleotide at the site of the labile nucleotide occurs by treatment with uracil N-glycosylase.
本发明公开了一种用于产生至少一种部分双链目标核酸的工艺,该核酸的末端含有至少一个单链区。该工艺包括以下步骤 (a) 提供至少一种引物 P
1
(b) 将至少一个靶核酸序列与 P
1
生成含有至少一个易损核苷酸的双链多核苷酸;(c) 将双链多核苷酸暴露于可促进多核苷酸在引物 P 1 的至少一个易损核苷酸位点处发生单链裂解的条件下。
1
和 (d) 将裂解的多核苷酸暴露于促进引物 P
1
从末端解离的条件。可变核苷酸可以是 dUTP,其中多核苷酸在可变核苷酸位点的单链裂解是通过尿
嘧啶 N-糖基化酶处理进行的。