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Home / Assay Kits

Human Mitochondrial RNA Polymerase Assay Kit

SKU : BHE16300024

This product includes all the assay kit components except the enzyme for 100 assays in a 384-well plate assay format: 400 ul of 10 x Buffer, 33 ul of 100 x DNA template, 33 ul of 100 x NTP mix, 330 ul of 10 x fluorescence dye. Enzyme is not included

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Mfr.No.	MRPA100K, MRPA100KE
Description	The human mitochondrial RNA polymerase (h-mtRNAP or POLRMT) functions as the transcriptase for expression and the primase for replication of mitochondrial DNA. It’s malfunction is related to various diseases and aging. Some antiviral nucleoside analogs were reported to cause chronic toxicity and related to inhibition of mitochondrial RNA polymerase activity. The Human Mitochondrial RNA Polymerase Assay is based on measurement of the RNA molecules synthesized by the RNA polymerase using a DNA template and DNTPs. It is fluorescence assay in a 384-well or 96-well plate format. The assay can be used for measurement activities of human mitochondrial RNA polymerase and drug screens against this enzyme. The Human Mitochondrial RNA Polymerase Assay Kit (Catalog No. MRPA100K) includes all the assay kit components except the enzyme for 100 assays in a 384-well plate assay format: 400 µl of 10 x Buffer, 33 µl of 100 x DNA template, 65 µl of 50 x NTP mix, 330 µl of 10 x fluorescence dye.
Drug target assays	Anticancer drug targets

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 50 x Human Mitochondrial RNA Polymerase 5-fold with the 1 x assay buffer. Each assay uses 3 µl of 10 x enzyme. (3) 10 x NTP mix: Dilute the 50 x NTP mix 5-fold with water. Each assay uses 3 µl of 10 x NTP mix. (4) 1 x dye: Dilute the 10 x fluorescence dye 10-fold with water. 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 2 hours. 3. Detection: Mix 30 µl of 1 x dye with 30 µl of the assay reaction mixture for 5 min and read the fluorescence at 535 nm (excitation at 485 nm).

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Combination SKU	Supplier No.	Specifications	Size	Your price	Quantity
BHE16300024-1	MRPA100K	enzyme not included	100 assays	$465.97		Add to Cart
BHE16300024-2	MRPA100KE	enzyme included	100 assays	$780.83		Add to Cart

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