Total RNA was purified from MSCs and NK cells using the RNeasy mini kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions and quantified by an average optical density (OD) OD260 nm/OD280 nm using a Synergy 2 Multi-Detection-Reader (BioTek). RNA was reverse transcribed using a SuperScript II RNase Reverse Transcriptase kit with Hexamer random primers (both Thermo Fisher Scientific), according to the manufacturer's protocol. Amplification was performed in a My-cycler thermal cycler (Bio-Rad, Munich, Germany). qRT-PCR was performed on diluted cDNA using DyNAmo Capillary SYBR Green qPCR kit (Thermo Fisher Scientific) and Roche LightCycler 2.0 instrument (Roche, Wiesbaden, Germany). Gene expression was quantified using the 2(−Δ ΔC(T)) method accordingly (Livak and Schmittgen, 2001 (link)). A list of primers and sequences is provided in Table S1 .
Dynamo capillary sybr green qpcr kit
Manufactured by Thermo Fisher Scientific
Sourced in Germany, United States
The DyNAmo Capillary SYBR Green qPCR kit is a real-time PCR reagent designed for quantitative PCR analysis using SYBR Green detection. The kit includes a master mix, containing a proprietary hot-start DNA polymerase, SYBR Green I dye, and necessary reaction components for efficient real-time PCR amplification and detection.
Automatically generated - may contain errors
Lab products found in correlation
Synergy 2 multidetection reader, by Agilent Technologies (1 mentions)
Hexamer random primers, by Thermo Fisher Scientific (1 mentions)
Lightcycler 2.0 instrument, by Roche (1 mentions)
Rneasy mini kit, by Qiagen (1 mentions)
Mycycler thermal cycler, by Bio-Rad (1 mentions)
Superscript 3 first strand synthesis system for rt qpcr, by Thermo Fisher Scientific (1 mentions)
Nucleospin rna, by Macherey-Nagel (1 mentions)
2 protocols using dynamo capillary sybr green qpcr kit
1
Quantitative RT-PCR of MSCs and NK Cells
2
Real-Time PCR Analysis of FFAR1 in Islet Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Total mRNA was isolated from islet cells using NucleoSpin® RNA (Macherey-Nagel, Duren, Germany) and reversely transcribed into cDNA with SuperScript™ III First-Strand Synthesis System for RT-qPCR (Invitrogen, Carlsbad, CA, USA). The real-time PCR was performed in 10 μL volume using Dynamo Capillary SYBR Green qPCR kit (Thermo Scientific, Waltham, MA, USA). The following primers were used for amplification: FFAR1 (forward primer, 5′-ATCACAGCCTTCTGCTAC and reverse primer, 5′-CCTAGATTGGGGTACAGG), β-actin (forward primer, 5′-ACGTGGACATCCGCAAAGAC) and (reverse primer, 5′-CAGGGCAGTGATCTCCTTCT). FFAR1 mRNA level was normalized to the reference gene β-actin using the following formula: target amount = 2−ΔΔCt, where ΔΔCt = [Ct (GPR40 KO) − Ct (β-actin KO)] − [Ct (GPR40 control) − Ct (β-actin control)]55 (link).
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