• Efficiently clone any insert of 35-45 kb.
  • Get more recombinants from less DNA.
  • High insert stability.
  • Inducible copy number.

Improved Fosmid Cloning

Lucigen’s CopyRight® v2.0 Fosmid Cloning Kit with the pSMART® FOS vector and Replicator™ FOS strain contains the most advanced fosmid cloning developed. The CopyRight v2.0 Fosmid Kit yields many more recombinants from limited starting DNA (Figure 1). The vector incorporates CopyRight transcription terminators to stabilize inserts, which results in complete fosmid libraries without missing clones. False-negatives and false-positives are dramatically reduced, and empty-vector background is essentially eliminated (Figure 2). Controllable genetic elements in the pSMART FOS vector (Figure 3) and Replicator FOS cells allow amplification of vector copy number for high plasmid yields (Figure 4).

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Figure 1. Higher recombinant yields with a limiting amount of DNA. Lucigen’s CopyRight Fosmid kit compared to Epicentre’s CopyControl kit.
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Figure 2. Zero background with the pSMART FOS vector (left) compared to the CopyControl™ pCC1FOS™ vector (right) (Epicentre).
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Figure 3. CopyRight v2.0 pSMART FOS vector. sacB, sucrase gene; lacZ, lacZ alpha peptide gene; Cm^r - chloramphenicol resistance gene; cosN - lambda packaging signal; par A,B,C - partition genes; ori2, repE [not shown in drawing], IncC - single-copy origin of replication; oriV - inducible origin of replication. Approximate positions of sequencing primers and transcription terminators (T) are indicated. The lacZ/sacB stuffer region is completely removed during vector processing.
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Figure 4. Copy number amplification of fosmid clones from the CopyRight v2.0 Fosmid Kit (Replicator cells) was more consistent and robust than the CopyControl Fosmid kit (EPI300 cells).

Convenient Success

Efficiently obtain your clone or create your library—the CopyRight v2.0 Cloning Kits eliminate tedious vector and competent cell preparation, as well as time-consuming QC testing experiments. Kits include optimized reagents, ligation-ready vector, optimized cells, detailed instructions, and trouble-shooting guide.

ORDER INFORMATION

Each CopyRight v2.0 Fosmid Cloning Kit contains: pre-cut, dephosphorylated pSMART FOS blunt vector, CloneSmart DNA Ligase, CloneDirect™ 10X Ligation Buffer (includes ATP), DNATerminator End Repair Enzyme Mix and End Repair Buffer, Replicator FOS strain (glycerol stock), 20% Maltose Solution, 1M MgSO4, SM Buffer, Arabinose Induction Solution, Sequencing Primers, and complete protocols. The 20-reaction Kit is provided as two 10-reaction Kits.