Verso sybr green 1 step qrt pcr low rox kit

Manufactured by Thermo Fisher Scientific

The Verso SYBR Green 1-Step qRT-PCR Low ROX Kit is a reagent kit designed for quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis. The kit utilizes SYBR Green dye for the detection and quantification of RNA targets.

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Lab products found in correlation

Tri reagent, by Merck Group (1 mentions) Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions) Nanodrop 8000, by Thermo Fisher Scientific (1 mentions) Quantitech primer assay, by Qiagen (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions)

Spelling variants of this product

SYBR Green (3216 citations) SYBR Green I (788 citations) SYBR Green kit (254 citations) QRT-PCR (101 citations) QRT-PCR kit (42 citations) SYBR Green qRT-PCR (21 citations) Verso (14 citations) SYBR I (3 citations)

2 protocols using verso sybr green 1 step qrt pcr low rox kit

RNA Extraction and qRT-PCR Analysis

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Total cellular RNA was isolated using the TRI-reagent (Sigma-Aldrich, USA, Cat. #T9424). Briefly, after respective treatments, cells were homogenized in TRI-reagent and total RNA was isolated as per the manufacturer’s protocol [19 (link)]. The quality and quantity of the isolated RNA were determined, and the RNA was aliquoted and stored at -80 °C until further analysis. qRT-PCR was performed using CFX96 Touch Real-Time PCR Detection System (BioRad Inc.). In brief, 50 ng of RNA template was used per reaction using the Thermo Scientific Verso SYBR Green 1-Step qRT-PCR Low ROX Kit (Cat. #AB-4106/C) as per the manufacturer’s protocol. GAPDH (glyceraldehyde-3-phosphate dehydrogenase) expression was utilized as a housekeeping control to quantify relative mRNA expression using the ΔΔCt method as previously described [19 (link)]. Primers used for Nrf-2 expression: Forward 5-CTGAACTCCTGGACGGGACTA-3’; Reverse 5’-CGGTGGGTCTCCGTAAATGG-3’ and for NF-κB expression: Forward 5’-AGCTGATGTGCATCGGCAAGTG-3’; Reverse 5’- GTAGCTGCATGGAGACTCGAACAG-3’.

Diwan B., Yadav R., Singh A., Kumar D, & Sharma R. (2023). Murine sterile fecal filtrate is a potent pharmacological agent that exerts age-independent immunomodulatory effects in RAW264.7 macrophages. BMC Complementary Medicine and Therapies, 23, 362.

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Quantifying Gene Expression in Stem Cells

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RNA was extracted from hiPSC and hESCs growing in a culture dish to 80 % of confluence, using RNeasy Plus Mini Kit (QIAGEN, 74134). RNA purity and concentration were confirmed using NanoDrop 8000 (Thermo Scientific).
RT-qPCR was performed on a Mastercycler ep RealPlex² Real Time PCR ThermoCycler (Eppendorf, 2894) using Verso SYBR Green 1-Step qRT-PCR Low ROX Kit (Thermo Scientific, AB-4106). Reactions were set up according to the manual using 1 ng RNA, 2.5 μL primers, 0.25 μL Verso Enzyme Mix, 1X SYBR mix, 1.25 μL RT Enhancer, and nuclease-free water for a total volume of 25 μL. The 1-Step RT-qPCR thermal cycling program was carried out at 50 °C for 15 min; 95 °C for 15 min; and 40 cycles of 95 °C for 15 s, 55 °C for 30 s and 72 °C for 30 s, with subsequent melting curve analysis. The primers used for RT-qPCR were Hs_FMR1_1_SG (QIAGEN QuantiTech^® Primer Assay, QT00017479) and Hs_GAPDH_1_SG (QIAGEN QuantiTech^® Primer Assay, QT00079247). Expression of transcripts of target genes was normalized to Gapdh.

Hiroi N., Brown J.R., Haile C.N., Ye H., Greenberg M.E, & Nestler E.J. (1997). FosB mutant mice: Loss of chronic cocaine induction of Fos-related proteins and heightened sensitivity to cocaine’s psychomotor and rewarding effects. Proceedings of the National Academy of Sciences of the United States of America, 94(19), 10397-10402.

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