Lucigen’s unique cloning kits with pre-processed vectors and highly competent cells eliminate hours of tedious preparation and enable efficient cloning. Ligation can be completed in as little as 30 minutes with no clean-up needed; only a brief heat denaturation is necessary before transformation. Downstream manipulation (e.g, mutagenesis, subsequent insert addition) is facilitated by the minimal vector size (1.7-2.0 kb).
All cloning kits include the transcription- and translation-free pSMART® vectors (Figure 1), which eliminate many of the problems associated with cloning recalcitrant DNA in conventional plasmid vectors. Conventional cloning vectors contain promoters (e.g., lacZ promoter) that constitutively transcribe the insert sequence. This transcription and coupled translation can destabilize inserts that contain coding regions, strong promoters, short repeats, or incompatible secondary structures. The cloning site of pSMART vectors does not include a lacz sequence. In addition, strong transcription terminators flank the cloning site to block spurious transcription from the vector and insert-driven transcription into the vector. Low copy number versions of pSMART further stabilize sequences that are difficult to maintain in typical vectors.
The pSMART vectors are supplied pre-digested, with blunt, dephosphorylated ends, and are qualified to produce 99.5% recombinant clones in typical experiments. The ultra-low background of empty vector (less than 0.5%) eliminates the need to screen for recombinants, removes the uncertainty of false negatives (light blue pUC colonies) and false positives (white colonies that lack inserts), and enables library construction from nanogram amounts of DNA.
Figure 1. High copy (HC) and low copy (LC) versions of the pSMART transcription-free cloning vectors.
Efficiently obtain your clone or create your genomic or cDNA library—the CloneSmart Cloning Kits eliminate tedious preparation of vectors and competent cells, as well as time-consuming QC testing experiments. Kits include optimized reagents, ligation-ready vector (no post-ligation cleanup step required), detailed instructions, and trouble-shooting guides to simplify cloning and sequencing. Highly efficient E. cloni® 10G electrocompetent cells (up to >4 × 1010 cfu/µg) or chemically competent cells ( >1 × 109 cfu/µg) are available for sale separately.
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