GC Cloning and Amplification (pGC™ Blue Vector)

Advanced PCR cloning
GC Cloning (patents pending) is the first major advance in PCR cloning since the first description of TA-cloning (Mead, D., et. al. (1991) Biotechnology 9, 657.) TA-cloning takes advantage of the well-known property of non-proofreading DNA polymerases (e.g., Taq, Tfl, Tth) to add a single 3´-A to PCR products.

The GC Cloning technology is based on the discovery that these same enzymes add a single 3´-G to DNA molecules, either during PCR or as a separate G-tailing reaction to any blunt DNA (Figure 1). The pGC™ Blue Vector (Figure 2) contains a single 3´-C overhang, which is compatible with the single 3´-G overhang on the inserts.  The pGC Blue vector includes the lacZ sequence for blue/white screening, as well as dual opposed RNA polymerase promoters for in vitro transcription of the cloned insert.

Stabilized inserts
The pGC Blue Vector incorporates the transcription--free CloneSmart technology (Figure 1) to eliminate many of the problems associated with cloning recalcitrant DNA [download eLucidations™ article (PDF) and PowerPoint presentation (PPT)].  Strong transcription terminators flank the cloning site to block spurious transcription from the vector into the insert and from the insert into the vector.

Convenient Success

Efficiently clone your PCR product regardless of its size, base composition, or enzyme used to generate it.  Kits include reagents for amplification and ligation, ligation-ready vector (no post-ligation cleanup step required), highly efficient electrocompetent cells (up to >4 X 10¹⁰ cfu/µg) or chemically competent cells ( >1 X10⁹ cfu/µg), detailed instructions, and trouble-shooting guides to simplify PCR cloning.