Quant-iT™ PicoGreen® dsDNA Assay Kit

Protocol tips

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	To minimize photo bleaching effects, keep the time for fluorescence measurement constant for all samples. During incubation at room temperature, protected from light.

Protocol tips
To minimize photo bleaching effects, keep the time for fluorescence measurement constant for all samples. During incubation at room temperature, protected from light.

Publication protocol

The yield of undigested DNA and purified mononucleosomal DNA was measured using Quant-iT™ PicoGreen™ dsDNA Assay Kit (Invitrogen, Carlsbad, CA, USA) in a Qubit™ 2.0 Fluorometer. DNA amounts obtained from various samples were normalized to make them equal. Equal amount of DNA was then used as template in qPCR in real time using the primers for α-globin, β-globin and pancreatic amylase loci. For each primer, percentage protection was calculated by comparing Ct values of undigested DNA versus mononucleosomal DNA for DMSO or 898 treatments and further normalized to Ct values of undigested DNA versus mononucleosomal DNA obtained from the serum starved cells.

Slot blot analysis of bromodeoxyuridine-labeled DNA
For measuring BrdU incorporation, NIH3T3 cells in S-phase were treated with DMSO or 3 μM Hdacs1,2-selective inhibitor (898) for 12 h, 18 h or 24 h followed by labeling of cells with 20 μM BrdU for 1 hr. Genomic DNA was isolated using Qiagen DNeasy™ Kit and extensively digested with RNaseA. DNA was quantified in a Qubit™ 2.0 Fluorometer using Quant-iT™ PicoGreen™ dsDNA Assay Kit (Invitrogen). Equal amount DNA (500 ng) from the different samples was resuspended in 40 μl water and denatured by adding 10 volumes of 0.4 N NaOH and incubation at room temperature for 30 min. Equal volume of 1 M Tris. Cl, pH 6.8 was added to neutralize and samples were placed on ice.

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Quant-iT™ PicoGreen® dsDNA Assay Kit below.

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