Cloning and Library Construction |
Fosmid, BAC, and Cosmid Cloning
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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.
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| 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. |
CopyControl™ Fosmid Library Production Kit
CopyControl™ HTP Fosmid Library Production Kit
EpiFOS™ Fosmid Library Production Kit
FosmidMAX™ DNA Purification Kit
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