pSCH2009

87419 ^™

This is a partial clone of the Klebsiella pneumoniae aac(6?)-Ib gene. Nucleotides 1-331 of the insert correspond to nucleotides 471 - 801 of the Genbank sequence M21682.A hybridization probe may be generated using the following vector specific PCR primers: Modified T3:5??CCCCTCACTAAAGGGAACAAAAGCTG ? 3?Modified T7:5? ? CGCGTAATACGACTCACTATAGGGCGAA-3?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. Due to the sequence similarity between aac(6?)-Ib and aac(6?)-IIa genes, the aac(6?)-Ib probe is used to detect both genes. - In: Woodford, N; Johnson, A., eds. Methods in molecular medicine: molecular approaches for the diagnosis and investigation of bacterial diseases. Totowa, NJ: The Humana Press, Inc.; 1996

Organism: Klebsiella pneumoniae (Schroeter) Trevisan
Clone type: Clone
Applications: Molecular biology
Product format: Freeze-dried
Shipping information: Escherichia coli containing the phagemid
Storage conditions: 2°C to 8°C

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Documentation

Product sheet

BSL 1

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

General

Specific applications: Due to sequence similarity between aac(6')-Ib and aac(6')-IIa genes, the aac(6')-Ib probe is used to detect both genes.

Characteristics

Comments: Restriction digests of the clone give the following sizes (kb): HindIII/XhoI--3.0, 0.4; HindIII--3.4; XhoI--3.4.
The insert contains the following restriction sites (approximate kb from the 5' end): ClaI--0.04; ScaI--0.11; SacI--0.32.
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.
Due to sequence similarity between aac(6')-Ib and aac(6')-IIa genes, the aac(6')-Ib probe is used to detect both genes.
A hybridization probe may be generated using the following vector specific PCR primers: modified T3 = 5'-CCCCTCACTAAAGGGAACAAAAGCTG-3' and modified T7 = 5'-CGCGTAATACGACTCACTATAGGGCGAA-3'.
Mycoplasma contamination: Not detected

Vector information

Construct size (kb): 3.299999952316284

Insert information

Insert size (kb): 0.33100000000000002
Type of DNA: cDNA
Gene product: aminoglycoside 6'-N-acetyltransferase [aac(6')-Ib]
Gene symbol: aac(6ʹ)-Ib

Handling information

Medium: ATCC Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Temperature: 37°C
Handling procedure: 1. Open vial according to instructions.2. aseptically add 0.3 to 0.4 mL of liquid medium to the freeze-dried pellet and mix well. Transfer 100 uL to a test tube containing 5 mL LB+ ampicillin (50-100 ug/mL). A loopful of culture can also be streaked on an agar plate of the same. Incubate cultures at 37⁰C. 3. Isolate DNA using standard plasmid preparation procedures.
Handling notes: Restriction digests of the clone gave the following sizes (in kb): HindIII/XhoI 3.0, 0.4 ; HindIII 3.4 ; XhoI 3.4. ATCC Staff

History

Depositors: KJ Shaw
Special collection: NCRR Contract
Cross references: GenBank M21682

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Intended use

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.

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Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

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References

Curated Citations

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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