pSCH2003 plasmid in E. coli (ATCC® 87427)

Organism: Unidentified bacterium  /  Clone Type: Clone  /  Depositors: KJ Shaw

Permits and Restrictions

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Designations pSCH2003 plasmid in E. coli
GenBank Number

X04555

Species Unidentified bacterium
Depositors KJ Shaw
Vector
Construct size (kb): 3.25
Insert
DNA: Synthetic
Insert lengths(kb): 0.305
Gene product: aminoglycoside adenylyltransferase(spectinomycin resistance, streptomycin resistance) [ant(2")-Ia]
Insert Size (kb) 0.305
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level 1
Comments
Restriction digests of the clone give the following sizes (kb): XhoI--3.3; HindIII--3.3; PstI--3.3; HindIII/XhoI--2.8, 0.4.
The insert contains the following restriction sites (approximate kb from the 5' end): PvuII--0.20; RsaI--0.25; SphI--0.29.
A single gel purification of the PCR generated probe is necessary since flanking regions will co-amplify with the gene specific sequence. Failure to do so often results in high backgrounds and false positives with clinical E. coli strains.
A hybridization probe may be generated using the following vector specific PCR primers: modified T3 = 5'-CCCCTCACTAAAGGGAACAAAAGCTG-3' and modified T7 = 5'-CGCGTAATACGACTCACTATAGGGCGAA-3'.
References

Shaw KJ, et alThe application of molecular techniques for the study of aminoglycoside resistanceIn: Shaw KJ, et alMethods in molecular medicine: molecular approaches for the diagnosis and investigation of bacterial diseasesTotowa, NJHumana Presssubmitted, 1996

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