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S. aureus RNA polymerase assay kit

BHE16300155

This product includes 400 ul 10 x Buffer, 33 ul 100 x DNA, 33 ul 100x NTPs, 350 ul 10 x fluorescence dye. The assay This product includes all reagents except the enzyme. It is for 100 assays of S. aureus RNA polymerase reactions in a 384-well assay format.

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+1 866.986.9598
Email
orders@biohippo.com
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Reference Siricilla S. et al, Discovery of a capuramycin analog that kills nonreplicating Mycobacterium tuberculosis and its synergistic effects with translocase I inhibitors. The Joual of Antibiotics, 68, 271–278 (2015).
Product Manual Datasheet
Protocols Description Assay Protocol
The following assay protocol is based on the 384-well plate assay format (plate type: Matrix 4318 or alike). The reaction volume is 30 µl and the final assay volume is 60 µl. For 96-well plate assays (plate type: Costar 3915 or alike), the reaction volume is 60 µl and the final assay volume is 120 µl.
 
1. Reagent preparation:
(1) 10 x DNA template: dilute the 100 x DNA template10-fold with water.
(2) 10 x enzyme: dilute the 100 x RNA polymerase 10-fold with the 1 x Buffer.
(3) 10 x NTP mix: dilute the 100 x NTP mix (50 mM) 10-fold with water.
(4) 1 x fluorescence dye: dilute the 10 x fluorescence dye 10-fold with water.
 
Note: The final concentration of S. aureus RNA polymerase in the assay is 20 nM. The assay buffer is 42.5 mM HEPES, pH 7.5, 42.5 mM NH4Cl, 2 mM DTT, 4 mM MgCl2, 0.005% Tween-20.
 
2. Reaction:
The total volume of each reaction mixture is 30 µl including 18 µl of H2O, 3 µl of 10 x Buffer, 3 µl of 10 x DNA template, 3 µl of 10 x enzyme, 3 µl of 10 x NTP mix. Incubate the reaction mixture in a standard black 384-well plate (Matrix 4318) at 37°C for 60 min.
 
3. Detection:
Add 30 µl of the 1 x fluorescence dye into the 30 µl of the reaction mixture. Incubate for 5 min. Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.
Combination SKU Supplier No. Specifications Size Your price Quantity
BHE16300155-1 RPA-100KS enzyme not included 100 assays $381.85
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BHE16300155-2 RPA-100KSE enzyme included 100 assays $640.73
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