Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM-F12), penicillin/streptomycin and fetal bovine serum (FBS) were obtained from Gibco. Collagenase P were obtained from Roche. Anti-human CD34-PE, CD45-FITC, HLA-DR-FITC, CD44-FITC, CD73-PE, CD90-FITC, and were purchased from BD Biosciences. Adipogenic and osteogenic inducing medium from Cyagen. Matrigel were obtained from Corning. Trizol Reagent were purchased by Invitrogen. All-In-One RT MasterMix Kit, EvaGreen qPCR MasterMix Kit were obtained from abmGood. Cell-tracker dye CM-DiI were purchased from Invitrogen.
Evagreen qpcr mastermix kit
Manufactured by Abmgood
Sourced in Canada
The EvaGreen qPCR MasterMix Kit is a laboratory reagent designed for quantitative polymerase chain reaction (qPCR) analysis. It contains the necessary components, including the EvaGreen DNA-binding dye, required to perform qPCR experiments.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (3 mentions)
All in one rt mastermix kit, by Abmgood (2 mentions)
All in one rt mastermix, by Abmgood (2 mentions)
Dulbecco s modified eagle s medium nutrient mixture f 12 dmem f 12, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Cd45 fitc, by BD (1 mentions)
Cd73 pe, by BD (1 mentions)
Hla dr fitc, by BD (1 mentions)
Cd44 fitc, by BD (1 mentions)
Cd90 fitc, by BD (1 mentions)
Anti human cd34 pe, by BD (1 mentions)
Matrigel, by Corning (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Collagenase p, by Roche (1 mentions)
Cell tracker dye cm dii, by Thermo Fisher Scientific (1 mentions)
Mirvana qrt pcr mirna detection kit, by Thermo Fisher Scientific (1 mentions)
Abi 7500 fast real time pcr system, by Thermo Fisher Scientific (1 mentions)
Abi 7500, by Thermo Fisher Scientific (1 mentions)
4 protocols using evagreen qpcr mastermix kit
1
Isolation and Characterization of Mesenchymal Stem Cells
2
Quantification of Cardiac Gene Expression
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Total RNA samples were extracted from cultured neonatal mice CMs or the left ventricle of mice hearts using the TRIZOL reagent (Invitrogen, Carlsbad, CA, USA). RNA was reverse-transcribed with 5X All-In-One RT MasterMix (abmGood, Vancouver, Canada). Real-time qPCR was carried out on an ABI 7500 fast real time PCR system (Applied Biosystems, Foster City, CA, USA) with the EvaGreen qPCR Mastermix Kit (abmGood, Vancouver, Canada) to detect the mRNA levels of Plscr4, ANP, BNP, and β-MHC, and GAPDH was set as the internal control. The miR-214 level was quantified by the mirVana qRT-PCR miRNA detection kit (Ambion, Austin, TX, USA) according to the manufacturer’s protocols and as previously described,44 (link), 45 (link) and these data were normalized to U6. The data were analyzed using the 2-ΔΔCT method. The sequences of the primers were synthesized by Sangon Biotech (Shanghai, China) and are listed in Table S2 .
3
Cardiac Fibrosis Quantification Protocol
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Total RNA were extracted from plasma or cardiac tissues from TAC mice. Total RNA was isolated from 1 ml plasma using phenol/chloroform extraction procedures. For qRT-PCR, the cDNA was assessed with 5X All-In-One RT MasterMix (abmGood, Canada). The EvaGreen qPCR Mastermix Kit (abmGood, Canada) was used in real-time PCR for relative quantification. Collagen, type I alpha1, Collagen type III alpha1, and serpinE2 mRNA levels were quantified by fast real-time PCR system (ABI 7500, Applied Biosystems, Carlsbad, CA, USA), and GAPDH was set as an internal control. The 2−ΔΔCt method was applied for the data analysis and the data were normalized and converted into relative mRNA expression.
4
Quantifying Gene Expression in hDPSCs
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Cultured hDPSCs in P4 were collected and total RNA were extracted with Trizol Reagent (Invitrogen). cDNA was synthesized from RNA using All-In-One RT MasterMix Kit (abmGood, Canada). qPCR was performed using EvaGreen qPCR MasterMix Kit (abmGood, Canada). Thermal cycling conditions were 95°C for 10 min and 40 cycles of 95°C for 15 s, 60°C for 60 s. The sequences of primers were listed in Table 1
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