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Description:
M13mp18 is the double-stranded, covalently closed, circular form of DNA derived from bacteriophage M13. This phage vector contains single HindIII, SphI, SbfI, PstI, SalI (AccI/ HincII), XbaI, BamHI, SmaI (XmaI), KpnI (Acc65I), SacI and EcoRI sites within the β-Galactosidase gene (1). When a fragment of DNA is inserted into one of these sites, the β-Galactosidase gene is inactivated, providing selection for clones on the appropriate indicator plate (2).
Source:
The phage M13mp18 is propagated in E. coli K12 JM101(3). The replicative form of DNA is isolated from infected cells and purified by gel filtration and column chromatography. The final preparation is tested for its suitability as a vector.
Concentration:
100 μg/ml
Storage Conditions
Storage Conditions:
10 mM Tris-HCl
1 mM EDTA
pH 7.5 @ 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.
Transformation Reaction::
Undigested vector (12 ng/100 µl competent cells) yielded 3.3 x 105 pfu/µg plaques. Of these, 100% were blue plaques and < 0.05% were colorless. EcoRI digested vector yielded 3% blue plaques and < 0.05% colorless plaques. EcoRI digested vector ligated in the absence of target DNA yielded 34% blue plaques and < 0.05% colorless plaques. EcoRI digested vector ligated in the presence of target DNA yielded 29% blue plaques and 7% colorless plaques.
References
- Norrander, J., Kempe, T. and Messing, J. (1983) Gene, 26, 101-106.
- Messing, J., Crea, R. and Seeburg, P.H. (1981) Nucl. Acids Res., 9, 309-321.
- Messing, J. (1979) Recombinant DNA Technical Bulletin (NIH), 2, 43-48.
Companion Products
M13mp18 Single-stranded DNA
M13mp19 RF I DNA
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