pWEB-TNC™ Cosmid Cloning Kit

pWEB-TNC™ Cosmid Cloning Kit


货号

WEBC931


规格

1 Kit


存储温度

MaxPlax is stored at -70℃, the remainder is stored at -20℃


产品详情


Applications

  • Preparation of complete, unbiased cosmid libraries.

The pWEB-TNC™ and the pWEB™ Cosmid Cloning Kits facilitate rapid and efficient construction of cosmid libraries using pWEB-TNC (Fig. 1) or pWEB (Fig. 2) Cosmid Vectors, respectively. The kits use the strategy of cloning end-repaired, randomly sheared DNA instead of the conventional approach of cloning fragments generated by partial restriction endonuclease digestion. 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™-T1R E. coli plating cells, all included in the kit. The result is a complete and unbiased primary cosmid library.

EPICENTRE also offers kits for fosmid cloning and library production. See related products for more information.

Benefits

  • pWEB cloning vectors are cloning-ready: linearized, dephosphorylated, and highly purified.
  • No need for partial restriction endonuclease digests or pulse-field gel electrophoresis to prepare the genomic DNA for cloning.
  • High-efficiency lambda packaging eliminates background and false positives.

 

Figure 1 (click to enlarge). Map and 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. Figure 2 (click to enlarge). Map and cloning region of the pWEB™ Vector. GenBank accession number AF075573. Figure 3 (click to enlarge). Outline of the cosmid cloning procedure using the pWEB-TNC™ and the pWEB™ Cosmid Cloning Kits.

 

Catalog No. Size

pWEB-TNC™ Cosmid Cloning Kit
WEBC931 1 Kit

For producing up to 10 complete and unbiased cosmid libraries.

Contents: pWEB-TNC™ Cosmid Vector, End-Repair Enzyme Mix, End-Repair 10X Buffer, dNTP Mix, Fast-Link™ DNA Ligase, Fast-Link™ 10X Ligation Buffer, ATP Solution, GELase™ Enzyme Preparation, GELase™ 50X Reaction Buffer, MaxPlax™ Lambda Packaging Extracts, Ligated Lambda Control DNA, Cosmid Control DNA, EPI100™-TR Plating Strain, Control Lambda Plating Strain, Ammonium Acetate Solution.

pWEB™ Cosmid Cloning Kit
PC8805 1 Kit

For producing up to 10 complete and unbiased cosmid libraries.

Contents: pWEB™ Cosmid Vector, End-Repair Enzyme Mix, End-Repair 10X Buffer, dNTP Mix, Fast-Link™ DNA Ligase, Fast-Link™ 10X Ligation Buffer, ATP Solution, GELase™ Enzyme Preparation, GELase™ 50X Reaction Buffer, MaxPlax™ Lambda Packaging Extracts, Ligated Lambda Control DNA, Cosmid Control DNA, EPI100™-TR Plating Strain, Control Lambda Plating Strain, Ammonium Acetate Solution.