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Home / Enzymes, Inhibitors & Substrates / Bacterial Enzymes

E. coli DNA primase - for 100 assays

BHE16300187

This product includes 45 ul of 10 uM E. coli primase-helicase complex

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+1 866.986.9598
Email
[email protected]
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Reference Lacriola CJ et al, Inhibition of DNA replication in Staphylococcus aureus by tegaserod. The Joual of Antibiotics. 70, 918–920 (2017). 
Product Manual Datasheet
Protocols Description Assay Protocol using the E. coli DNA Primase Assay Kit
1. Reagent preparation:
10 x DNA: dilute the 100 x DNA with water.
10 x enzyme: Dilute the 100 x enzyme stock with the 1 x assay buffer to make the 10 x enzyme.
10 x NTP mix: dilute the 100 x NTP 10-fold with water.
1 x fluorescence dye: dilute the 10 x fluorescence dye 10-fold with water.
 
2. Reaction:
The total volume of each reaction mixture is 40µl including 24µl of H2O, 4µl of 10 x Buffer, 4µl of 10 x DNA template, 4µl of 10 x enzyme, 4µl of 10 x NTP mix. Incubate the reaction mixture at 37°C for 60 min.
 
Note: The assay solution is composed of 10 mM HEPES, pH 7.5, 5 mM magnesium acetate, 0.5 mM DTT, 0.003% Brij-35, 100 nM DNA, 0.5 mM NTPs, 100 nM enzyme.
 
3. Detection:
Add 80µl of the 1 x fluorescence dye into the 40µl of the reaction mixture. Incubate for 5 min. Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.
SKU Supplier No. Size Your price Quantity
BHE16300187 DNAG-100EC 100 assays $436.31
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