pWEB™Cosmid Cloning Kit

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Last modified: 2008-07-15 14:57:15

Everything you need for producing complete and unbiased cosmid libraries in about 2 days

The pWEB-TNC™ and the pWEB™ Cosmid Cloning Kits facilitate rapid and efficient construction of cosmid libraries. Using pWEB-TNC™ (Figure 1) or pWEB™ Cosmid Vectors, respectively. The kits utilise 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 linearised and dephosphorylated pWEB-TNC™ or pWEB™ Cosmid Vector, packaged using ultra-high efficiency MaxPlax™ Lambda Packaging Extracts (>10⁹pfu/µg for phage lambda), also included in the kit, and plated on the supplied EPI100™-T1^R E. coli, plating cells. The result is a complete and unbiased primary cosmid library.

Epicentre® offers two kits for cosmid cloning and library production. Each kit contains complete instructions and all reagents needed for production of up to 10 cosmid libraries.

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 the 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

Protocols for: pWEB™ Cosmid Cloning Kits

(catalogue number WEBC931 / TNC9401)

(catalogue number PC8805)

(catalogue number TNC1SP01)

Please note: all protocols off site are the responsibility of the products supplier

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