Triple-Ligation Strategy with Advantages Over Directional Cloning

Triple-ligation strategy with advantages over directional cloning.

Useful properties of restriction enzymes that recognize interrupted palindromes.

ed here improves upon the concept of directional cloning without forfeiting any of its advantages. The advantages include single-step cloning, high-efficiency generation of the desired recombinant molecule and low background self-ligation of vector. The added benefits of the triple-ligation strategy include easy resolution of vector fragments for gel purification, the ability to invert an inser...

Quick and Clean Cloning: A Ligation-Independent Cloning Strategy for Selective Cloning of Specific PCR Products from Non-Specific Mixes

We have developed an efficient strategy for cloning of PCR products that contain an unknown region flanked by a known sequence. As with ligation-independent cloning, the strategy is based on homology between sequences present in both the vector and the insert. However, in contrast to ligation-independent cloning, the cloning vector has homology with only one of the two primers used for amplific...

Recursive directional ligation by plasmid reconstruction allows rapid and seamless cloning of oligomeric genes.

This paper reports a new strategy, recursive directional ligation by plasmid reconstruction (PRe-RDL), to rapidly clone highly repetitive polypeptides of any sequence and specified length over a large range of molecular weights. In a single cycle of PRe-RDL, two halves of a parent plasmid, each containing a copy of an oligomer, are ligated together, thereby dimerizing the oligomer and reconstit...

Synthesis of homogeneous MUC1 oligomers via a bi-directional ligation strategy.

The efficient synthesis of homogeneous MUC1 peptide oligomers using sequential ligation reactions in the N-to-C and C-to-N directions is reported. The bi-directional ligation strategy makes use of thioester formation via N → S acyl shift chemistry in combination with peptide ligation reactions and was used to prepare a library of peptide oligomers ranging in molecular mass from 3.8-9.4 kDa, com...