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T7 RNA Polymerase Assay Kit

BHE16300033

This product includes 400 ul of 10 x Buffer, 33 ul of 100 x DNA, 33 ul of 100 x NTPs, 1550 ul of 2 x Dye and 1550 ul of 50 mM EDTA. It is for 100 assays of T7 RNA polymerase. The assay This product includes all reagents except the enzyme.

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Tel
+1 866.986.9598
Email
[email protected]
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Ask Merchant
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Reference Siegmund V. et al, Screening mutant libraries of T7 RNA polymerase for candidates with increased acceptance of 2'-modified nucleotides, Chem. Commun., 48, 9870-9872 (2012).
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: Dilute the 100 x DNA 10-fold with water. Each assay uses 3 µl of 10 x DNA.
(2) 10 x enzyme: Dilute the 100 x T7 RNA polymerase (50 Unit / µl) 10-fold with the 1 x assay buffer. Each assay uses 3 µl of 10 x enzyme.
(3) 10 x NTP mix: Dilute the 100 x NTP mix (50 mM) 10-fold with water. Each assay uses 3 µl of 10 x NTP mix.
(4) 1 x dye: Dilute the 2 x fluorescence dye 2-fold with 50 mM EDTA. Each assay uses 30 µl of 1 x dye.
 
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 at 37°C for 60 min.
 
3. Detection:
Mix 30 µl of the 1 x fluorescence dye with 30 µl of the reaction mixture. Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.
Combination SKU Supplier No. Specifications Your price Quantity
BHE16300033-1 T7RPA100K enzyme not included $348
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BHE16300033-2 T7RPA100KE enzyme included $465.97
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