A hybridization-based approach for quantitative and low-bias single-stranded DNA ligation

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review

33 Scopus Citations
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Author(s)

  • Chun Kit Kwok
  • Yiliang Ding
  • Madeline E. Sherlock
  • Sarah M. Assmann
  • Philip C. Bevilacqua

Detail(s)

Original languageEnglish
Pages (from-to)181-186
Journal / PublicationAnalytical Biochemistry
Volume435
Issue number2
Publication statusPublished - 2013
Externally publishedYes

Abstract

Single-stranded DNA (ssDNA) ligation is a crucial step in many biochemical assays. Efficient ways of carrying out this reaction, however, are lacking. We show here that existing ssDNA ligation methods suffer from slow kinetics, poor yield, and severe nucleotide preference. To resolve these issues, we introduce a hybridization-based strategy that provides efficient and low-bias ligation of ssDNA. Our method uses a hairpin DNA to hybridize to any incoming acceptor ssDNA with low bias, with ligation of these strands mediated by T4 DNA ligase. This technique potentially can be applied in protocols that require ligation of ssDNA, including ligation-mediated polymerase chain reaction (LMPCR) and complementary DNA (cDNA) library construction. © 2013 Elsevier Inc. All rights reserved.

Research Area(s)

  • Hybridization, Ligation, Low-sequence bias, Nucleotide preference, Single-stranded DNA

Citation Format(s)

A hybridization-based approach for quantitative and low-bias single-stranded DNA ligation. / Kwok, Chun Kit; Ding, Yiliang; Sherlock, Madeline E. et al.
In: Analytical Biochemistry, Vol. 435, No. 2, 2013, p. 181-186.

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review