Construction of new T vectors for direct cloning of PCR products

Gene. 1993 Aug 16;130(1):153-4. doi: 10.1016/0378-1119(93)90361-6.

Abstract

More than half of the products of PCR contain an extra A residue at the 3' end, which is the result of the template-independent activity of Taq polymerase. To facilitate cloning of the products of PCR without modification, T vectors, which have a single overhanging T residue at the 3' end, have been developed. In the present study, we constructed new T vectors which can be prepared in the laboratory by simple digestion with the restriction enzymes AspEI or Eam1 105I.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular / methods*
  • Deoxyribonucleases, Type II Site-Specific
  • Genetic Vectors*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Polymerase Chain Reaction*
  • Thymine

Substances

  • endodeoxyribonuclease Eam1105I
  • Deoxyribonucleases, Type II Site-Specific
  • Thymine