EZ-Tn5™ Transposase

pWEB::TNC™ Cosmid Cloning Kit

For Research Use Only. EZ-Tn⁵™ products covered by issued and pending patents

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Last modified: 2008-12-10 15:08:44

The pWEB-TNC™ and the pWEB™ Cosmid Cloning Kits facilitate rapid and efficient construction of cosmid libraries using pWEB-TNC™ (Figure 1) or pWEB™ (Figure 2) Cosmid Vectors, respectively. The kits utilize a novel strategy (Figure 3) of cloning end-repaired, randomly sheared DNA instead of the conventional approach of cloning fragments generated by partial restriction endonuclease digestion. First, genomic DNA is sheared by passing it through a syringe needle (not supplied with either kit). The sheared DNA is end-repaired to generate 5'-phosphorylated blunt ends and size-selected using a low melting point agarose gel. The size-selected DNA is then ligated into the supplied linearized and dephosphorylated pWEB-TNC™ or pWEB™ Cosmid Vector, packaged using ultra-high efficiency MaxPlax™ Lambda Packaging Extracts (>10⁹ pfu/µg for phage lambda) and plated on phage T1-resistant EPI100™-T1^R E. coli plating cells, all included in the kit. The result is a complete and unbiased primary cosmid library.

Epicentre® also offers kits for cosmid cloning and library production. See related products for more information.

pWEB-TNC™ Cosmid Cloning Kit

The pWEB-TNC™ Cosmid Cloning Kit contains the pWEB-TNC™ Cosmid Vector. This vector can be used in all the same ways as the original pWEB™ vector, but offers additional features that can be used if and when desired. For example, any of the cosmid clones produced in the pWEB-TNC™ vector can be used for subsequent generation of a random unidirectional deletion library from any chosen cosmid clone when used in conjunction with the pWEB-TNC™ Deletion Cosmid Transposition Kit.

pWEB™ Cosmid Cloning Kit

The pWEB Cosmid Cloning Kit is EPICENTRE's® original kit for rapid and easy cosmid cloning and library production.

Figure 1. Map & cloning region of the pWEB-TNC™ Vector.The Sma I cloning site is flanked by pairs of BamH I, EcoR I, and Not I sites to aid in the excision and mapping of insert DNA. If desired, unidirectional deletion subclones can be generated from any clone in pWEB-TNC using the pWEB-TNC Deletion Cosmid Transposition Kit.	Figure 2. Map and cloning region of the pWEB™ Vector.GenBank accession number AF075573.

Figure 3. Outline of the cosmid cloning procedure using the pWEB-TNC™ and the pWEB™ Cosmid Cloning Kits.

Last modified: 2008-05-14 16:46:46

For Research Use Only. EZ-Tn⁵™ products covered by issued and pending patents

Get your Epicentre® Forum - hot off the press! Click here for direct link to Forum listings

Last modified: 2008-12-10 15:08:44