CopyControl cDNA, Gene and PCR Cloning

Features & Benefits

• Single-copy clone growth ensures clone stability
• Inducible high-copy amplification for high DNA yield
• Ideal for cloning of unstable inserts and cDNAs encoding toxic proteins
• Lower risk of mutated clones
• Better representation of libraries

Description

Up to ~25% more cDNA clones can be obtained in a library if the cDNA clones are present in the host cell at only about one copy per cell. This is presumably due to lower detrimental effects on host cell metabolism and lower toxicity of expressed clone sequences and gene products.

The CopyControl cDNA, Gene, and PCR Cloning Kits provide a rapid method for cloning any blunt-end DNA fragments at single-copy number, with the capability to induce to high-copy numbers as needed.

The CopyControl pCC1 (Blunt) Vector provided in the kits contains both the E. coli F-factor single-copy origin of replication (oriII) and an inducible high-copy oriV. Replication does not occur using the oriV unless and until the trfA gene in the chromosome of TransforMax™ EPI300™ cells (which is under the control of a tightly regulated promoter) is induced.

Thus, CopyControl cDNA, PCR, or gene clones can be grown at single copy to ensure insert stability and successful cloning of sequences that are toxic to the host cell. Then, any desired clone can be induced up to 200 copies per cell for high yields of DNA for all downstream applications.