Construct fosmid libraries of 40 kb clones with high efficiency using improved systems in standard or high-throughput format.

• Prepare unbiased fosmid libraries with a complete kit, including GELase enzyme and MaxPlax Lambda Packaging Extracts
• Stabilize large or difficult fosmid clones with tight control of copy number
• Optimize high-throughput end-sequencing results with CopyControl HTP Fosmid Library Production Kit

Improve your fosmid libraries with CopyControl™ Fosmid Library Production Kits

The CopyControl Fosmid Library Production Kit uses the pCC1FOS™ Vector, which contains both the E. coli F-factor single-copy origin of replication and the inducible high-copy oriV. The oriV replication origin allows CopyControl Fosmid clones to be grown at single copy, to ensure insert stability and successful cloning of toxic and unstable DNA sequences. Immediately before DNA purification, CopyControl Fosmid clones can be induced up to 50 copies per cell to greatly increases DNA yields while maintaining the plasmid stability.

The CopyControl HTP Fosmid Library contains the pCC2FOS™ Vector which is designed for high-throughput settings to optimize end-sequencing results. The primer cassette, engineered in conjunction with Agencourt Bioscience Corporation, eliminates wasteful and expensive vector sequence reads by having the 3´ends of the primer-annealing sites only three bases from the vector/insert junction. In addition, the seven-base sequence at the 3´end of each primer was specifically designed to minimize mispriming to any contaminating E. coli DNA present after template purification.

The kit uses a streamlined cloning protocol. First, shear genomic DNA into approximately 40-kb fragments, then end-repair the sheared DNA to generate blunt, 5´-phosphorylated ends. This produces a more complete and unbiased genomic library than can be obtained by partial restriction endonuclease digests. After DNA size selection and recovery, ligate the DNA fragments into the cloning-ready CopyControl pCC1FOS or pCC2FOS Vector, package with the included ultra-high efficiency MaxPlax™ Lambda Packaging Extracts (>109 pfu/µg DNA), and plate on the supplied TransforMax™ EPI300™ E. coli. Once the library is prepared, individual clones can be cultured in small volumes and induced to multiple-copy number for high yields of high-purity DNA for fingerprinting, sequencing, or other applications.

Eliminate background and false positives with high-efficiency lambda packaging, and use the easy "hands-off" autoinduction protocol for high-throughput applications. Fosmid cloning is faster and easier than BAC cloning, using simplified protocols and improved vectors.