Evagreen digital pcr supermix

Manufactured by Bio-Rad

EvaGreen Digital PCR Supermix is a ready-to-use reagent designed for digital PCR applications. It contains all the necessary components, including the EvaGreen dye, for performing digital PCR experiments.

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Lab products found in correlation

Maxima h minus cdna synthesis master mix, by Thermo Fisher Scientific (1 mentions) Quantasoft analysis pro software, by Bio-Rad (1 mentions) Quantasoft version 1, by Bio-Rad (1 mentions) Qx200 droplet reader, by Bio-Rad (1 mentions) Qx200 droplet digital pcr system, by Bio-Rad (1 mentions) Droplet generation oil for evagreen, by Bio-Rad (1 mentions) Qx200 droplet digital pcr ddpcr system, by Bio-Rad (1 mentions) Gotaq qpcr master mix, by Promega (1 mentions) Qscript cdna supermix, by Quantabio (1 mentions) Qs5 system, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

EvaGreen Supermix (113 citations) EvaGreen (51 citations) QX200 EvaGreen Digital PCR Supermix (4 citations) PCR Supermix (2 citations)

3 protocols using evagreen digital pcr supermix

Quantitative Gene Expression Analysis of Placental Transcripts

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RNA was extracted from placenta as described above. A two-step reverse transcriptase ddPCR (RT-ddPCR) protocol was used to quantify transcript abundance (46 (link)). cDNA was synthesized from total RNA using the Maxima™ H Minus cDNA Synthesis Master Mix (ThermoFisher). PCR primers targeting guinea pig transcripts were designed using the Primer Quest tool and were ordered from Integrated DNA Technologies (Table S2). ddPCR reactions were prepared using the EvaGreen Digital PCR Supermix (Bio-Rad) using a primer concentration of 250 nM and cycled using the following thermal conditions: 95°C for 5 min; 40 cycles of 95°C for 30 s and 60°C for 30s; 4°C for 5 min; 90°C for 5 min; hold at 4°C. The ddPCR data were analyzed with QuantaSoft™ Analysis Pro software (Bio-Rad). Absolute quantification of gene expression was presented as copies per nanogram of total RNA.

Berkebile Z.W., Putri D.S., Abrahante J.E., Seelig D.M., Schleiss M.R, & Bierle C.J. (2021). The Placental Response to Guinea Pig Cytomegalovirus Depends Upon the Timing of Maternal Infection. Frontiers in Immunology, 12, 686415.

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Quantification of miRs in Plasma-Derived MVs

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The expression of miRs in plasma‐derived MVs was quantified by means of the QX200 TM Droplet Digital TM PCR System (ddPCR, Bio‐Rad) using EvaGreen Digital PCR Supermix (Bio‐Rad) with Droplet Generation Oil for EvaGreen (Bio‐Rad) following manufacturer's instruction. Briefly, 20 μL of EvaGreen Supermix, 2 μM primers, and pre‐amplified cDNA were transferred in a droplet generation cartridge and mixed with 70 μL of droplet generation oil to obtain an emulsion. Droplets (40 μL) were transferred to a new ddPCR 96‐well plate. Amplification of specific miRs was obtained with the following protocol: 95°C 5 min 1 cycle enzyme activation, 95°C 30 s 40 cycles denaturation, 54°C 1 min 40 cycles annealing/extension, 4°C 5 min 1 cycle signal stabilization, 90°C 5 min 1 cycle signal stabilization, 4°C 40 min 1 cycle, 4°C. Fluorescence of single droplets was read using QX200 Droplet Reader, and data were analysed using the QuantaSoft Version 1.7.4 Software (Bio‐Rad) and expressed in copy number per microlitre.

Pin F., Beltrà M., Garcia‐Castillo L., Pardini B., Birolo G., Matullo G., Penna F., Guttridge D, & Costelli P. (2021). Extracellular vesicles derived from tumour cells as a trigger of energy crisis in the skeletal muscle. Journal of Cachexia, Sarcopenia and Muscle, 13(1), 481-494.

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Reverse Transcription and qPCR Analysis

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One microgram of RNA was reverse transcribed using Qscript cDNA Supermix (QuantaBio). qPCR on cDNA or ChIP DNA was performed with GoTaq qPCR Master Mix (Promega) under the manufacturer’s instruction on a QS5 system (Thermo Scientific). The copy numbers were measured by predetermined standard curves. The relative expression or enrichment to control samples was calculated based on the ΔΔCt method. Quantitative digital droplet PCR was performed on a QX200 Droplet Digital PCR (ddPCR) System (Bio-Rad) with EvaGreen Digital PCR Supermix (Bio-Rad) under the manufacturer’s instructions. Primers used in this study are listed in Supplementary Data 1.

Teo W.W., Cao X., Wu C.S., Tan H.K., Zhou Q., Gao C., Vanuytsel K., Kumar S.S., Murphy G.J., Yang H., Chai L, & Tenen D.G. (2022). Non-coding RNA LEVER sequestration of PRC2 can mediate long range gene regulation. Communications Biology, 5, 343.

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