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USER Friendly Cloning Vectors >
pNEB206A Linearized Vector |
 | | | | pNEB206A Linearized Vector | | | |
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Prices are in US dollars and valid only for US orders.
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Description:
Linearized pNEB206A Vector is supplied with the USER™ Friendly Cloning Kit. pNEB206A has been linearized within the Multiple Cloning Site to produce 8-nucleotide, 3´ single-stranded extensions on both vector ends. The single-stranded extensions are not complementary; this prevents the vector termini from re-annealing to form transformable circular DNA and also allows control over the orientation of the inserted PCR product. pNEB206A linearized vector is 2,706 base pairs in length.
Source:
pNEB206A is isolated from E. coli by a standard purification procedure, digested to completion with XbaI and nicked with Nt.BbvCI. The DNA is phenol extracted and resuspended in 10 mM Tris-HCl (pH 8.0), 1 mM EDTA
Concentration:
20 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 8.0 @ 25°C
Storage Temperature:
-20°C
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
A standard USER reaction:
A standard USER reaction was performed as described in Appendix V of the USER Friendly Cloning Kit manual [20 ng linearized pNEB206A, 1 µl USER Enzyme and 10 µl (100 ng) of a 950 bp control PCR product amplified using Taq DNA Polymerase and primers containing uracil, designed as recommended in the USER Friendly cloning Kit manual]. After transformation into chemically-competent cells (NEB #ER2267 at 5 x 106 c.f.u./µg pNEB206A), 50 µl of the 1 ml outgrowth was spread on Amp + Xgal + IPTG plates. A minimum of 200 colonies were obtained and > 95% of these were white (i.e., contained recombinant molecules).
Companion Products
USER Enzyme
USER Friendly Cloning Kit
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