ERP Micro December 2019

1)

Amplification (Amp) tubes Tq, containing lyophilized

(l) Micropipettor tips.— Filter-barrier, volumes 20–200 and 200–1000 µL. (m) Incubator.— Maintaining 42 ± 1°C. (x) Masticator. —Seward 400 (Seward Ltd, Worthington, UK; www.seward.co.uk), or equivalent. Optional : The following apparatus are required for use with Assurance® GDS for E . coli O157:H7 Tq assay (576 tests, Cat. No. 71007-576) due to the reduced size of the Amplification tubes. (a) Amplification tube holder – (Cat. No. 79134BC), variable spacing, 72-well (b) Amplification tube holder lid – (Cat. No. 79184BC), variable spacing, 72-well (c) Amplification tube capping tool – (Cat. No. 79132BC) (d) Amplification tube cap rack – (Cat. No. 79197BC), 72-well (e) Aluminum cooling block – (Cat. No. 73093BC), 72-well (f) Assurance GDS Rotor-Gene® Q Rotor-Disc – (Cat. No. 73076BC), 72-well (g) Assurance GDS Rotor-Gene® Q locking ring (Cat. No. 73084BC), 72-well, to lock the Rotor-Disc For information on additional materials needed for sample analysis by the Assurance® GDS PickPen® PIPETMAX® (PPMX) please see the PPMX User Manual (No. 55240BC). (n) C. Media and Reagents Item ( a ) is available from BioControl Systems. Items ( b )–( e ) are available in the Assurance GDS E. coli O157:H7 Tq test kit from BioControl Systems. Item ( f ) is available commercially. Items are available from MilliporeSigma (www.sigmaaldrich.com). Ordering information—SigmaAldrich, 12822 SE 32nd St, Bellevue, WA 98005, Tel: 425-586-3300, email: Prewarm 1 L sterile deionized water at 42 ± 1˚C overnight. On day of use aseptically transfer 31.6 g mEHEC® media into the prewarmed sterile water. Gently mix to fully rehydrate the media. Use prepared broth within 6 h. Alternatively, mEHEC® broth can be prepared and autoclaved at 121˚C for 15 min. Autoclaved broth must be prewarmed to 42 ± 1˚C overnight prior to sample addition. (b) Assurance® GDS for E . coli O157:H7 Tq Assay – (100 tests; Cat. No. 71007-100), kit includes bcs_cust_svc_bellevue@milliporesigma.com. (a) modified EHEC broth (mEHEC®)— Optional : Assurance® GDS PickPenTM PipetMax® (Cat. No. 79100BC)

reagents for E . coli O157:H7, 1 pouch, Cat. No. 34739– 100 2)

O157 Concentration Reagent—One bottle, Cat. No.

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34030 – 100 3)

Resuspension Buffer Tq—One bottle, Cat. No. 34724 –

100

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4) Wash Solution—One bottle, Cat. No. 34043 – 100 (c) Assurance® GDS for E . coli O157:H7 Tq Assay – (576 tests; Cat. No. 71007-576), kit includes 1) Amplification (Amp) tubes Tq, containing lyophilized reagents for E . coli O157:H7, 1 pouch, Cat. No. 34739– 576 2) O157 Concentration Reagent—Two bottles, Cat. No. 34030 – 576 3) Resuspension buffer Tq—Two bottles, Cat. No. 34724 – 576 4) Wash solution—Three bottles, Cat. No. 34043 – 576 (d) Assurance® GDS for E . coli O157:H7 Tq Assay – (576 tests; Cat. No. 71007-576ATM), for use with automated (PipetMax) sample preparation, kit includes 1) Amplification (Amp) tubes Tq, containing lyophilized reagents E . coli O157:H7, 4 pouches, Cat. No. 34749-144 2) O157 Concentration Reagent—Two bottles, Cat. No. 34030 – 576 The following reagents are also necessary but sold separately: 3) Resuspension buffer Tq— Cat. No. 34724-100C 4) Wash solution— Cat. No. 61031-100 (a) BioControl mEHEC broth .—Prewarm 1 L sterile deionized water at 42 ± 1°C overnight. On day of use, aseptically transfer 31.6 g mEHEC into the prewarmed sterile water. Gently mix to dissolve thepowder.Usepreparedmediumwithin6 h.Alternatively, mEHEC can be prepared and autoclaved at 121°C for 15 min. Autoclaved media must be prewarmed to 42 ± 1°C overnight prior to sample addition. (b) Concentration reagent . (c) Resuspension buffer . (d) Amplification tubes containing lyophilized reagents for E. coli O157:H7 . (e) Wash solution. (f) Diagnostic media and reagents .—Necessary for cultural confirmation of positive Assurance GDS samples. Reagents ( b ) and ( d ) must be stored at 2–8°C when not in use. D. General Instructions Reagents (1) through (4) must be stored at 5 ± 3°C when not in use.

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(a) Store Assurance® GDS for E . coli O157:H7 Tq kit components at 5 ± 3°C. Kit expiration is provided on the product box label. Do not use Assurance GDS for E E . coli O157:H7 Tq reagents that have expired. The quality of expired materials is not warrantied. (b) Read the kit instructions carefully before use. Failure to do

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