title: A hybridization-based approach for quantitative and low-bias single-stranded DNA ligation.
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.
authors: Department of Chemistry, The Pennsylvania State University, University Park, PA 16802, USA.
authors: Chun Kit
authors: CK
authors: Kwok
authors: Yiliang
authors: Y
authors: Ding
authors: Madeline E
authors: ME
authors: Sherlock
authors: Sarah M
authors: SM
authors: Assmann
authors: Philip C
authors: PC
authors: Bevilacqua
mesh_terms: D001483:Base Sequence; D011088:DNA Ligases / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000032:analysis*; D015723:Gene Library; D055029:Inverted Repeat Sequences; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction
chemical_list: D004277:DNA, Single-Stranded; D011088:DNA Ligases