H9812沙门氏菌标准菌株-DNA转用于脉冲场凝胶电泳分子量标准 BioVector NTCC质粒载体菌种细胞基因保藏中心

H9812沙门氏菌标准菌株-DNA转用于脉冲场凝胶电泳分子量标准

BioVector NTCC质粒载体菌种细胞基因保藏中心

Salmonella enterica subsp.

enterica serovar

Braenderup
Designation: H9812 [MIS00418]

Deposited Name: Salmonella choleraesuis subsp. choleraesuis (Smith) Weldin serotype Braenderup

Antigenic Properties: I 6,7:e,h:e,n,z15

Product Description: The DNA from this strain is used by laboratories in the Pulsenet system as the

molecular size standard for pulsedfield

gel electrophoresis (PFGE) analysis of bacterial pathogens (see

notes).

Medium

Trypticase Soy Agar/Broth

Growth Conditions

Temperature: 37°C

Atmosphere: Aerobic

Propagation Procedure

1. Open vial according to enclosed instructions.

2. Using a single tube of #18 broth (5 to 6 mL), withdraw approximately 0.5 to 1.0 mL with a Pasteur or

1.0 mL pipette. Rehydrate the entire pellet.

3. Aseptically transfer this aliquot back into the broth tube. Mix well.

4. Use several drops of the suspension to inoculate a #18 slant, and/or plate.

5. Incubate all tubes and plate at 37°C. Growth should occur in 24 hours.
Instructions for Testing BAA664

Using the PulseNet Standardized PulsedField

Gel

Electrophoresis (PFGE) Protocols

If you have ordered this strain for the purpose of conducting PulsedField

Gel Electrophoresis (PFGE) analysis

using any of the standardized PulseNet protocols, please read the following information.

PFGE plugs (or blocks) of the Salmonella serotype Braenderup H9812 strain  are made

according the “PulseNet OneDay

(2428

h) Standardized Laboratory Protocol for Molecular Subtyping of E.

coli O157:H7, Salmonella serotypes, and Shigella sonnei by PFGE.” A copy of this protocol can be

requested by sending an Email

to pfge@cdc.gov.

This strain is used as a size standard for the normalization and analysis of PFGE patterns for all organisms

tracked by PulseNet, including E. coli O157:H7, Salmonella, Shigella, Listeria monocytogenes, a n d

Campylobacter jejuni. After plugs of the size standard are made, approximately 2mm

slices are cut and

restricted with 4050

Units of XbaI enzyme for 2 hours at 37°C. The plug slices are loaded on the

electrophoresis gel in lanes 1, 5, 10 (10well

gel), 1, 5, 10, 15 (15well

gel), or 1, 5, 10, 15, 20 (20well

gel).

Test samples restricted with the appropriate enzyme(s) are loaded in the other lanes. Electrophoresis is done

using the appropriate conditions for the test organism. The table on the next page shows the PFGE pattern

that is obtained with different electrophoresis conditions. The electrophoresis run time may have to be

optimized in each laboratory so that the lowest band in the H9812 standard migrates 1.0 1.5

cm from the

bottom of the gel. New lots of S. Braenderup H9812 PFGE plugs should be tested with “old” lots to confirm

that the pattern and band intensity is the same and that no observable genetic changes have occurred.