Vilo master mix kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The VILO Master Mix kit is a reagent formulation designed for reverse transcription of RNA into cDNA. It enables efficient and reliable conversion of RNA to cDNA for downstream applications such as PCR and qPCR analysis.

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Lab products found in correlation

Rnaspin mini rna isolation kit, by GE Healthcare (2 mentions) Abi prism 7000 device, by Thermo Fisher Scientific (2 mentions) Nanodrop nd 1000, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Sybr green pcr master mix, by New England Biolabs (1 mentions)

Close spelling variants of this product

VILO kit (25 citations) VILO master mix (23 citations)

3 protocols using vilo master mix kit

Quantitative PCR Analysis of Collagen Gene

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The quantitative PCR technique was performed following the Minimum Information for Publication of Quantitative Real-Time PCR Experiments guidelines (Bustin et al. 2009 (link)). Total ribonucleic acid (RNA) (i.e. 3 μg) was isolated from liver tissue using the RNAspin mini RNA isolation kit (GE HealthCare, USA) and was reverse transcribed to copy DNA using random primers and VILO Master Mix kit (Invitrogen). Collagen α1(I) primers and probes (assay ID Mm00801666_g1) for real-time PCR were purchased from Applied Biosystems (USA). 18S rRNA (assay ID Mm04277571_s1) and ACTB (assay ID Mm00607939_s1) were used as reference genes to normalize the results. Each sample was analyzed in duplicate and negative controls were enrolled. Efficiency was verified and established between 95% and 105%. Analyses were carried out using an ABI PRISM 7000 device (Applied Biosystems, USA). Analyses of relative gene expression data were performed according to the 2^-ΔΔCq method (Livak and Schmittgen, 2001 (link)). Results were expressed as fold change of ΔΔCq values obtained from WT oil mice.

Cogliati B., Crespo Yanguas S., da Silva T.C., Aloia T.P., Nogueira M.S., Real-Lima M.A., Chaible L.M., Sanches D.S., Willebrords J., Maes M., Pereira I.V., de Castro I.A., Vinken M, & Dagli M.L. (2016). Connexin32 deficiency exacerbates carbon tetrachloride-induced hepatocellular injury and liver fibrosis in mice. Toxicology mechanisms and methods, 26(5), 362-370.

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Quantitative PCR Analysis of Skin Tissue

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The qPCR technique was performed following the MIQE guidelines [17 (link)]. Total RNA (i.e. 3 μg) was isolated from skin tissue using the RNAspin mini RNA isolation kit (GE HealthCare, USA) and was reverse transcribed to cDNA using random primers and VILO Master Mix kit (Invitrogen). Primers and probes assays for real-time PCR were purchased from Applied Biosystems (USA), including those for: collagen type I (assay ID Mm00801666_g1), collagen type III (assay ID Mm00802332_m1), transforming growth factor beta 1 (TGFβ-1; assay ID Mm00441724_m1), matrix metallopeptidase 2 (MMP-2; assay ID Mm00439508_m1). 18S rRNA (assay ID Mm04277571_s1) and ACTB (assay ID Mm00607939_s1) were used as reference gene to normalize the results. Each sample was analyzed in duplicate and negative controls were enrolled; its efficiency was verified and established between 95% and 105%. Analyses were carried out using an ABI PRISM 7000 device (Applied Biosystems, USA). Analyses of relative gene expression data were performed according to the 2^−ΔΔCq method [18 (link)]. Results were expressed as fold change of ΔΔCq values obtained from WT mice at the respective day of measurement.

Cogliati B., Vinken M., Silva T.C., Araújo C.M., Aloia T.P., Chaible L.M., Mori C.M, & Dagli M.L. (2015). Connexin 43 deficiency accelerates skin wound healing and extracellular matrix remodeling in mice. Journal of dermatological science, 79(1), 50-56.

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Quantifying Osteoblast Gene Expression

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RNA was extracted from MC3T3-E1 cells using Trizol reagent (Invitrogen, Life Technology) as following the manufacturer's protocols. RNA concentration and RNA purity were quantified by using NanoDrop (NanoDrop ND-1000, ThermoScientific). cDNA synthesized using superscript VILO Mastermix kit (Invitrogen, Thermo Fisher Scientific, Netherlands). The composition of cDNA synthesis contains 1 μg of RNA, 4 μL of master mix VILO, and RNase-free water to a final volume of 20 μL. The reaction followed the procedure by the manufacturer's guidance. The primers for marker genes using in qRT-PCR were runx2, osx, opn, col1, and housekeeping gene gapdh as an internal control. The list of test genes and the primer sequences were shown in Table 1. Quantitative PCR (Thermo Fisher Scientific, US) was performed using SYBR green PCR master mix (NEB, US) and analyzed by relative quantification method. T1-5 Table 1: Primer sequences of study genes.

Nguyen P., Nguyen H., Nguyen M., Nguyen H., Pham Q., & Ha N.X. (2021). Ethyl acetate extract of Smilax glabra Roxb roots and its major active compound astilbin promote osteoblastogenesis in vitro by upregulating bone cell differentiation- associated genes. Asian Pacific Journal of Tropical Biomedicine, 11(12), 553-553.

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