CloneMiner™ II cDNA Library Construction Kit

The CloneMiner™ II cDNA Library Construction Kit is a second generation CloneMiner™ kit that enables rapid construction of highly representative cDNA libraries without restriction enzyme cloning. This innovative library construction technology combines SuperScript™ III Reverse Transcriptase with Gateway™ cloning technology, resulting in the discovery of previously unobtainable, full-length clones. The kit also avoids the use of time-consuming, inefficient ligation reactions, making library construction faster and offering better representation.

The CloneMiner™ II cDNA Library Construction Kit ensures:
• High primary titers
• Large average insert sizes
• The highest percentage of full-length genes
• Highly efficient cloning of cDNA to multiple destination vectors without the need of restriction enzyme digestion and ligation

How the CloneMiner™ II Kit Works:

• High yields of full-length cDNA: The CloneMiner™ II kit contains SuperScript™ III for generating high cDNA yields. SuperScript™ III Reverse Transcriptase (RT), a proprietary mutant of SuperScript™ II RT, is active at 50°C and has a half-life of 220 minutes. It also has a point mutation that reduces RNase H activity, thereby decreasing RNA degradation during first-strand synthesis and increasing the percentage of full-length genes.

• Gateway™ cloning technology avoids restriction enzyme cloning: Library construction is mediated by Gateway™ Technology, a site-specific recombination system that eliminates the use of restriction enzymes and ligase in cloning. Each cDNA insert is flanked by specific att recombination sites (added during the cDNA synthesis steps) that recombine with complementary att sites present in Gateway™ donor vectors to create entry clones. Entry clones are subsequently recombined with Gateway™ expression vectors to create expression clones, effectively replacing the use of restriction enzymes and ligase. The resulting clones maintain the original orientation and reading frame enabling functional analysis of full-length genes and whole libraries.

How is This Kit Different From the Original CloneMiner™ Kit?
• The new kit incorporates a simplified protocol.
• To ensure high cDNA yields, SuperScript™ II Reverse Transcriptase (RT) has been replaced by SuperScript™ III Reverse Transcriptase.
• To enable reaction setup with fewer pipetting steps, Gateway™ BP Clonase™ Enzyme Mix has been replaced by Gateway™ BP Clonase™ II Enzyme Mix, which contains enzymes and buffer in a single mix.