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

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

Research output: Contribution to journalArticlepeer-review

21 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)181-186
Number of pages6
JournalAnalytical Biochemistry
Volume435
Issue number2
DOIs
StatePublished - 2013

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'A hybridization-based approach for quantitative and low-bias single-stranded DNA ligation'. Together they form a unique fingerprint.

Cite this