Introduction
The Xi-Clone PCR cloning method saves hours of work because it eliminates the need for restriction or ligation enzymes. PCR-amplified genes are simply mixed with the linearized Xi-Clone vectors and transformed into high efficiency DH5alpha competent E. coli. Greater than 85% of the resultant colonies will be recombinant clones that contain the insert in the correct orientation for expression.
Because the Xi-Clone vectors contain no multiple cloning sites or enzyme recognition sites (e.g. topoisomerase I recognition site), no unwanted sequences are incorporated into the resulting expression vector. This is especially important when it is necessary to minimize the structural or functional interference caused by unwanted sequences.
The Xi-Clone vectors are available for either mammalian expression or in vitro transcription/translation. The phCMV Xi-Clone vectors contain a highly optimized human cytomegalovirus (CMV) promoter together with an efficient SV40 terminator for superior expression levels in mammalian cells. The pIX Xi-Clone vectors contain a T7 promoter and T7 terminator for optimal expression in in vitro transcription/translation systems. these vectors also feature the option of native protein expression or epitope-tagged protein expression.
Each Xi-Clone PCR Cloning Kit may be purchased optionally with 250 μl of SmartCells Competent E. coli. These specially optimized competent cells provide transformation efficiencies of >1x109 cfu/μg, even under the most difficult transformation conditions (such as when more DNA is used for transformation, diluted DNA samples are used, or when a high salt ligation mix is used). SmartCells Competent E. coli are also available separately.
Contents
Xi-Clone PCR Cloning Vector (1 vial, 24 ìl)
Xi-Clone CAT Control Template (1 vial, 10 μl)
SmartCells Competent E. coli (5 vials x 50 μl) (Optional - extra charge)*
*
Please note that SmartCells Competent E. coli are now offered either separately or together with any Xi-Clone phCMV (XMC101020) or pIX Kit (XIC101020).
Storage
The Xi-Clone Cloning kit is shipped frozen. For maximum stability and long-term use, store SmartCells Competent E. coli at -70ºC and store all other reagents at -20 ºC upon receipt. All components are stable for 6 months if stored properly.
How the XiCloneTM PCR Cloning Kit Works
