TY - JOUR
T1 - A hybridization-based approach for quantitative and low-bias single-stranded DNA ligation
AU - Kwok, Chun Kit
AU - Ding, Yiliang
AU - Sherlock, Madeline E.
AU - Assmann, Sarah M.
AU - Bevilacqua, Philip C.
N1 - Funding Information:
This work was supported by Human Frontier Science Program (HFSP) grant RGP0002/2009-C to P.C.B. and S.M.A. and by a Penn State Summer Discovery grant to M.E.S. We thank S. Tan for the laboratory-made T4 DNA ligase.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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.
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U2 - 10.1016/j.ab.2013.01.008
DO - 10.1016/j.ab.2013.01.008
M3 - Article
C2 - 23399535
AN - SCOPUS:84874069421
SN - 0003-2697
VL - 435
SP - 181
EP - 186
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 2
ER -