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Home / Assay Kits

Human DNA Polymerase Alpha

BHE16300226

This product includes 7 ul of 4 uM human DNA polymerase alpha catalytic subunit.

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Product Manual Datasheet
Protocols Description Assay Protocol using Human DNA Polymerase Alpha Assay kit (Catalog No. HDPA100K)
The following assay protocol is based on the 384-well plate assay format (plate type: Matrix 4318 or alike). The reaction volume is 30µl and the final assay volume is 60µl. For 96-well plate assays (plate type: Costar 3915 or alike), the reaction volume is 60µl and the final assay volume is 120µl.
 
1. Reagent preparation:
(1) 10 x DNA: Dilute the 100 x DNA 10-fold with water. Each assay uses 3µl of 10 x DNA.
(2) 10 x enzyme: The recommended final enzyme concentration is 4 to 8 nM in the assay.
(3) 10 x dNTP: Dilute the 100 x dNTP (10 mM) 10-fold with water. Each assay uses 3µl of 10 x dNTP.
(4) 1 x dye: Dilute the 2 x fluorescence dye 2-fold with 50 mM EDTA. Each assay uses 30µl of 1 x dye.
 
2. Reaction:
The total volume of each reaction mixture is 30µl including 18µl of H2O, 3µl of 10 x Buffer, 3µl of 10 x DNA template, 3µl of 10 x enzyme, 3µl of 10 x dNTP. Incubate the reaction mixture at 37°C for 60 min. Note: The enzyme reaction rate will be slower if the assay is at a lower temperature.
 
3. Detection:
30µl of the 1 x fluorescence dye with 30µl of the reaction mixture for 5 min. Measure the fluorescence intensity at 535 nm using the excitation wavelength at 485 nm.
Combination SKU Supplier No. Size Your price Quantity
BHE16300226-1 HDPA1-100 100 assays $486.75
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BHE16300226-2 HDPA1-1000 1000 assays $2123.68
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