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E. coli DNA polymerase alpha - for 100 assays

BHE16300184

This product includes 33 ul of 2 uM E. coli DNA polymerase III alpha.

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+1 866.986.9598
Email
[email protected]
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Product Manual Datasheet
Protocols Description Assay Protocol using the DNA Polymerase III Alpha Assay Kit
1. Reagent preparation:
10 x DNA: dilute the 100 x DNA with water.
10 x enzyme: prepare 100 x DNA polymerase using the 1 x buffer for dilution.
10 x dNTP mix: dilute the 100 x dNTP mix 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 30µl including 18µl of H2O, 3µl of 10 x Buffer DP, 3µl of 10 x DNA, 3µl of 10 x enzyme, 3µl of 10 x dNTP mix. Incubate the reaction mixture at 37°C for 60 min.
 
Note: The final concentrations are 20 mM HEPES, pH 7.5, 10 mM Mg(OAc)2 , 1 mM DTT, 30 nM DNA, 20 nM DNA polymerase III alpha subunit, 0.1 mM dATP, 0.1 mM dGTP.
 
3. Detection:
Add 30µl of the 1 x fluorescence dye into the 30µl 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
BHE16300184 DPA-100EC 100 assays $421.56
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