Rapid and scalable in vitro production of single-stranded DNA-Reference-Cited by-同舟云学术

Rapid and scalable in vitro production of single-stranded DNA

Published:2019-02-22 Issue: Volume: Page:
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Author:

Minev Dionis,Guerra Richard,Kishi Jocelyn Y.^ORCID,Smith Cory,Krieg Elisha,Said Khaled,Hornick Amanda,Sasaki Hiroshi M.,Filsinger Gabriel,Beliveau Brian J.,Yin Peng,Church George M.,Shih William M.

Abstract

We present a rapid, scalable, user-friendly method for in vitro production of high-purity single-stranded DNA (ssDNA) ranging from 89–3315 nucleotides in length. PCR with a forward primer bearing a methanol-responsive polymer generates a tagged amplicon that enables selective precipitation of the modified strand under denaturing conditions. We demonstrate that the recovered ssDNA can be used for CRISPR/Cas9 homology-directed repair in human cells, DNA-origami folding, and fluorescent in situ hybridization.

Publisher

Cold Spring Harbor Laboratory

Reference20 articles.

1. Efficient preparation of single-stranded DNA for in vitro selection

2. [29] Rapid production of single-stranded sequencing template from amplified DNA using magnetic beads

3. Scalable amplification of strand subsets from chip-synthesized oligonucleotide libraries

4. In vitro synthesis of gene-length single-stranded DNA

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