A Look at PCR Cloning Tools

In 1972, two groups, working independently, developed a new method to clone fragments of DNA using the enzyme terminal deoxynucleotidyl transferase (TdT). Because the use of restriction enzymes for cloning purposes had not yet caught on, these two groups had developed their methodologies using enzymes and technologies that were readily available to them at the time. David Jackson, working in Paul Berg’s lab and Peter Lobban and A.D. Kaiser, used TdT to generate short deoxyadenylate and deoxythymidylate tails on linearized plasmid and “foreign” DNA. The two DNA’s were then mixed and incubated with exonuclease III, DNA Pol I, to fill in any gaps, and ligase, to covalently seal the recombinant molecule.

Almost 20 years later, this method for cloning would be modified and adapted to clone PCR products. Because taq DNA Pol preferentially adds adenine residues to the 3’ ends of PCR products, it has become common to use linearized vectors containing complementary overhanging thymidine residues to clone them. Fortunately, the exonucleases and DNA polymerases are no longer required for these cloning vectors. And in some cases, even the ligases can be left out. Take a look below at what’s available in PCR cloning tools.

Featured Products

Tools
Invitrogen
TOPO® Tools Technology provides a fast, flexible, and customizable method to build linear constructs for use in a wide range of molecular biology applications such as in vitro transcription, transient expression in mammalian cells, or mammalian two-hybrid analysis. Simply choose the promoter, tag, or other sequence elements you want and add them to the 5´ and 3´ ends of your PCR-amplified product to create a linear construct that can be used in your downstream applications. You'll get results without traditional cloning methods or passage through E. coli, saving 2-3 days of research time.
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PCR Library Construction System
Stratagene
The PCR Library Construction kit combines polymerase chain reaction (PCR) for amplifying cDNA and ligation-independent cloning (LIC) for directionally cloning PCR products to produce high-efficiency cDNA libraries in a plasmid vector. This methodology allows you to start with enriched poly(A)+ RNA ) or as little as 100 ng of total RNA
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QIAGEN PCR Cloning System
QIAGEN
The pDrive Cloning Vector provides highly efficient cloning of PCR products through UA hybridization. The vector is supplied in a linear form with a U overhang at each 3' end, which hybridizes with high specificity to the A overhang of PCR products generated by Taq and other non-proofreading DNA polymerases. The Ligation Master Mix, supplied in a convenient premixed format, is specifically designed to provide optimal hybridization conditions for efficient cloning.
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