Methods for processing and amplifying nucleic acids

摘要

The present invention is directed generally to the use of stem-loop oligonucleotides comprising an inverted repeat, a loop, and at least one degradable site or site capable of becoming a degradable site in the preparation of DNA molecules, such as a library. In particular embodiments, the invention employs a single reaction mixture and conditions. In particular, at least part of the inverted palindrome is removed during the preparation of the molecules to facilitate amplification of the molecules. Thus, in specific embodiments, the DNA molecules are suitable for amplification and are not hindered by the presence of the palindrome.

主权项

1. A method of producing a primer for use in an amplification reaction comprising:
(a) providing at least one stem-loop oligonucleotide comprising an inverted repeat, a loop, and at least one degradable site or site capable of becoming a degradable site; (b) subjecting the at least one stem-loop oligonucleotide to conditions such that a phosphodiester bond of the at least one stem-loop oligonucleotide at the at least one degradable site or site capable of becoming a degradable site is broken, thereby producing primers for use in an amplification reaction; and (c) carrying out an amplification reaction using the primers.