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  • Biotechnological production of ssDNA with DNA-hydrolyzing deoxyribozymes.

Biotechnological production of ssDNA with DNA-hydrolyzing deoxyribozymes.

STAR protocols (2021-05-25)
Jin Liu, Hongzhou Gu
ABSTRACT

Preparation of long single-stranded (ss)DNA in large quantities with high efficiency and purity remains a synthetic challenge. Here, we present a protocol for using DNA-hydrolyzing DNA enzymes (deoxyribozymes) for efficient biotechnological production of milligrams of ssDNA with a customizable sequence up to a few kilobases. Our protocol provides a convenient yet economical way to store the sequence information of target ssDNA on phages for selective mass production on demand. For complete details on the use and execution of this protocol, please refer to Jia et al. (2021).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Lithium chloride, for molecular biology, ≥99%
Sigma-Aldrich
Poly(ethylene glycol), BioUltra, 8,000
Sigma-Aldrich
Sodium acetate, anhydrous, for molecular biology, ≥99%
Sigma-Aldrich
Acrylamide, suitable for electrophoresis, ≥99%
Sigma-Aldrich
Bromophenol Blue, titration: suitable
Sigma-Aldrich
N,N′-Methylenebis(acrylamide), 99%
Sigma-Aldrich
Sodium dodecyl sulfate, BioReagent, suitable for electrophoresis, for molecular biology, ≥98.5% (GC)