Rapid and scalable in vitro production of single-stranded DNA

Minev, D.; Guerra, R.; Kishi, J.; Smith, C.; Krieg, E.; Said, K.; Hornick, A.; Sasaki, H.; Filsinger, G.; Beliveau, B.; Yin, P.; Church, G.; Shih, W.

doi:10.1101/558429

bioRxiv · 10.1101/558429

Rapid and scalable in vitro production of single-stranded DNA

Minev, D.·Guerra, R.·Kishi, J.·Smith, C.·Krieg, E.·Said, K.·Hornick, A.·Sasaki, H.·Filsinger, G.·Beliveau, B.·Yin, P.·Church, G.·Shih, W.

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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.

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Minev, D., Guerra, R., Kishi, J., Smith, C., Krieg, E., Said, K., Hornick, A., Sasaki, H., Filsinger, G., Beliveau, B., Yin, P., Church, G., Shih, W.. 2019-02-22. Rapid and scalable in vitro production of single-stranded DNA. https://doi.org/10.1101/558429

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