QuantiFluor® dsDNA System

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Exposure to light will reduce the sensitivity of the assay. Store QuantiFluor® dsDNA Dye and working solution protected from light

Upstream tips
Exposure to light will reduce the sensitivity of the assay. Store QuantiFluor® dsDNA Dye and working solution protected from light

Publication protocol

The allelic ratios of mutations in each sample used in rounds 2 and 3 were accurately quantified by a commercial sponsor (Horizon Diagnostics, Cambridge, UK) using droplet digital PCR (ddPCR) on a BioRad QX100 (Hercules, CA, USA) platform. Genomic DNA (gDNA) was extracted from FFPE sections on the Promega (Madison, WI, USA) Maxwell System using the Maxwell 16 FFPE Plus LEV DNA purification kit, according to the manufacturer’s protocol. Quantification was performed using a Promega QuantiFluor dsDNA assay kit, according to the manufacturer’s protocol. ddPCR was performed using Taqman custom SNP 40 × primer/probe assays (Life Technologies, Carlsbad, CA, USA) to assess the frequency of each mutation with the exception of the p.(E746_A750) assay, which was designed in-house. DNA (40 ng) was added to each ddPCR reaction. Reactions were performed in quadruplicate and droplets were generated using a Droplet Generator according to the manufacturer’s instructions. PCR was performed on a standard thermocycler using previously optimised, assay-specific cycling conditions. Droplets were analysed using a QX100 Droplet Reader as described in the manufacturer’s instructions. Data from at least 45 000 useable droplets were collected for each sample. Formalin-fixed paraffin-embedded reference standards (Horizon Diagnostics) were included as assay controls.

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Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA quantification Colorectal aenocarenoma (SW48) - paraffin embeded using QuantiFluor® dsDNA System from Promega.

Manufacturer protocol

Download the product protocol from Promega for QuantiFluor® dsDNA System below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA quantification Colorectal aenocarenoma (SW48) - paraffin embeded using QuantiFluor® dsDNA System from Promega. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.