One step plus system

Manufactured by Thermo Fisher Scientific

Sourced in United States

The One Step Plus system is a compact real-time PCR instrument designed for quantitative gene expression analysis. The system features a thermal block that can accommodate a range of sample formats, enabling flexible experimental workflows.

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Lab products found in correlation

Quantiscript, by Qiagen (3 mentions) Taqman probe, by Thermo Fisher Scientific (2 mentions) Superscript 3, by Thermo Fisher Scientific (2 mentions) Rneasy plus kit, by Thermo Fisher Scientific (2 mentions) Midi prep kits, by Qiagen (2 mentions) Rneasy ffpe kit, by Qiagen (1 mentions) Superscript 3 reverse transcriptase, by Thermo Fisher Scientific (1 mentions) Rnalater, by Qiagen (1 mentions) Rneasy plus mini kit, by Qiagen (1 mentions) Sybr green master mix, by Thermo Fisher Scientific (1 mentions) Iscript cdna synthesis kit, by Bio-Rad (1 mentions) Rneasy kit, by Qiagen (1 mentions) Trizol, by Thermo Fisher Scientific (1 mentions) Rever tra ace α transcriptase, by Toyobo (1 mentions) Random hexamer, by Thermo Fisher Scientific (1 mentions) Midiprep kit, by Qiagen (1 mentions) Taqman assay, by Thermo Fisher Scientific (1 mentions) Superscript vilo master mix, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

PLUSTM (3 citations) Step One andStep One Plus Real time PCR systems (3 citations) Step One Plus™ Real-Time PCR System Thermal Cycler (3 citations) Step One Plus sequence amplification system (3 citations) One Step One Plus™ System (2 citations) Step One Plus or 7500 Fast systems (2 citations) Real-time PCR Systems Step One Plus (2 citations) Applied Bio-systems Step One Plus RT-PCR system (2 citations) QPCR Step-One Plus detection system (2 citations) Step One Plus real-time PCR (RT-PCR) system (2 citations)

9 protocols using one step plus system

Gene Expression Analysis via qPCR

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mRNA was isolated (Qiagen RNeasy Plus Kit) and transcribed to cDNA (Invitrogen SuperScript III). Gene expression was analyzed using Taqman probes and the OneStep Plus system (Applied Biosystems). Each biological sample was analyzed in experimental replicate (n=2 repeated wells of the qPCR reaction) and the Ct value of each replicate was averaged and handed as n=1 unique biologic sample. Expression for each sample was normalized to β-Actin (ACTA1) gene expression (ΔCt) and relative to peripheral lung tissue control samples (ΔΔCt), and fold change calculated by 2-ΔΔCt (24 (link)). A total of n=3 unique biological samples were analyzed for each experiment.

Gilpin S.E., Li Q., Evangelista-Leite D., Ren X., Reinhardt D.P., Frey B.L, & Ott H.C. (2017). Fibrillin-2 and Tenascin-C Bridge the Age Gap in Lung Epithelial Regeneration. Biomaterials, 140, 212-219.

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Quantifying mRNA Expression Levels

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RNA was isolated using QIAGEN Midi-Prep Kits and RT with Quantiscript (QIAGEN) using random hexamers (Invitrogen). mRNA levels were measured with specific primers (Table S2) using SYBR green on a One Step Plus system (Applied Biosystems). Relative levels of each target gene were calculated using the DDCt formula and 18S RNA as a control.

Reardon C.A., Lingaraju A., Schoenfelt K.Q., Zhou G., Cui C., Jacobs-El H., Babenko I., Hoofnagle A., Czyz D., Shuman H., Vaisar T, & Becker L. (2018). Obesity and Insulin Resistance Promote Atherosclerosis through an IFNγ-Regulated Macrophage Protein Network. Cell reports, 23(10), 3021-3030.

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Isolation and Expression Analysis of RNA from Formalin-Fixed Paraffin-Embedded Bowel Tissue

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RNA was isolated from tissue that had been fixed in 4% paraformaldehyde in PBS overnight and embedded in paraffin blocks. For both in vitro cultured and transplanted bowel, three unique pieces of tissue each were compared, with samples taken a minimum of 0.5 cm in distance apart. Each sample consisted of four 15 μm sections that were deparaffinized with xylene. mRNA was isolated using the RNeasy FFPE Kit (Qiagen) and transcribed to cDNA (Invitrogen SuperScript III). Gene expression was analyzed using Taqman probes and the OneStep Plus system (Applied Biosystems). Each sample was analyzed in 2 replicate qPCR reactions, with the Ct value of each replicate averaged and treated as an n = 1 unique biologic sample. Gene expression for each sample was normalized to human β-Actin (ACTA1) expression (ΔCt) and relative to human jejunum control samples (DDCt), with fold change calculated by 2-ΔΔCt^{37 (link)}.

Kitano K., Schwartz D.M., Zhou H., Gilpin S.E., Wojtkiewicz G.R., Ren X., Sommer C.A., Capilla A.V., Mathisen D.J., Goldstein A.M., Mostoslavsky G, & Ott H.C. (2017). Bioengineering of functional human induced pluripotent stem cell-derived intestinal grafts. Nature Communications, 8, 765.

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RNA Isolation and Gene Expression Analysis

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Samples were stored in RNALater (Qiagen) until RNA isolation by RNeasy-Plus Mini Kit (Qiagen). RNA was transcribed to cDNA by SuperScript^® III Reverse Transcriptase (Life Technologies). Gene expression was quantified by Taqman^® assays using the OneStepPlus system (Applied Biosystems). Gene expression was analysed by ΔΔCt method with normalization to 18S gene expression.

Gilpin S.E., Ren X., Okamoto T., Guyette J.P., Mou H., Rajagopal J., Mathisen D.J., Vacanti J.P, & Ott H.C. (2014). Enhanced Lung Epithelial Specification of Human iPSCs on Decellularized Lung Matrix. The Annals of thoracic surgery, 98(5), 1721-1729.

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Quantifying TRF2 and miR-23a Levels

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Real-time qPCR for TRF2 was carried out as described previously (Wang et al., 2013 (link)). Briefly, total RNAs were isolated using QIAGEN RNeasy Kit (Qiagen, Valencia, CA, USA) and reverse-transcribed using the iScript cDNA Synthesis Kit (Bio-Rad, Hercules, CA, USA). GAPDH was used as internal control for normalization. For miR-23a, total RNAs were isolated using Trizol (Invitrogen, Carlsbad, CA, USA) and reverse-transcribed using ReverTra-Ace-α-Transcriptase (TOYOBO, Japan). U6 small nuclear RNAs were used as an internal control for normalization. All qPCRs were performed using SYBR green master mix (Applied Biosystems, USA) on Applied Biosystems One-step-plus system.
The primers for miR-23a and U6 were purchased from Ribobio, Guangzhou, China.
The TRF2 primers are TRF2-forward (5′- CCCAAGAACAAGCGCATGAC-3′) and TRF2-reverse (5′-GGGTTGGTTGAGAACGGTGG-3′). The GAPDH primers are GAPDH-forward (5′-GGAGCGAGATCCCTCCAAAAT-3′) and GAPDH-reverse (5′-GGCTGTTGTCATACTTCTCATGG-3′).
The TRF2 3′UTR qPCR primers for RNA pull-down are TRF2 3′UTR-forward (5′- CCAGGTTGATGACAGACCAG-3′) and TRF2 3′UTR-reverse (5′-AGATGTTGACAGCAAATGCC-3′).

Luo Z., Feng X., Wang H., Xu W., Zhao Y., Ma W., Jiang S., Liu D., Huang J, & Songyang Z. (2015). Mir-23a induces telomere dysfunction and cellular senescence by inhibiting TRF2 expression. Aging Cell, 14(3), 391-399.

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Quantifying mRNA Expression Levels

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RNA Isolation and qRT-PCR Analysis

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RNA was isolated using Qiagen Midi-prep kits and reverse-transcribed with Quantiscript (Qiagen) using random hexamers (Invitrogen), and mRNA levels were measured with specific primers (Table S1) using SYBR green on a One Step Plus system (Applied Biosystems). Relative levels of each target gene were calculated using the ΔΔCt formula, using 18S RNA as a control.

Tiwari P., Blank A., Cui C., Schoenfelt K.Q., Zhou G., Xu Y., Khramtsova G., Olopade F., Shah A.M., Khan S.A., Rosner M.R, & Becker L. (2019). Metabolically activated adipose tissue macrophages link obesity to triple-negative breast cancer. The Journal of Experimental Medicine, 216(6), 1345-1358.

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Quantifying Gene Expression in Lung Tissue

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mRNA was isolated (Qiagen RNeasy Plus Kit) and transcribed to cDNA (Invitrogen SuperScript III). Gene expression was analyzed using Taqman probes and the OneStep Plus system (Applied Biosystems). Each biological sample was analyzed in experimental replicate (n=2 repeated wells of the qPCR reaction) with the Ct value of each replicate averaged and handed as n=1 unique biologic sample. Expression for each sample was normalized to β-Actin (ACTA1) gene expression (ΔCt) and relative to cadaveric peripheral lung tissue control samples (ΔΔCt), with fold change calculated by 2^−ΔΔCt (33 (link)).

Gilpin S.E., Charest J.M., Ren X., Tapias L.F., Wu T., Da Silva D.E., Mathisen D.J, & Ott H.C. (2016). Regenerative Potential of Human Airway Stem Cells in Lung Epithelial Engineering. Biomaterials, 108, 111-119.

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Gene Expression Quantification in Alveolar Spheres

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RNA was isolated by Trizol then reverse transcribed to cDNA by SuperScript Vilo master mix (Life Technologies). Gene expression was quantified by Taqman assay (Life Technologies) using the One Step Plus system (Applied Biosystems). Gene expression was analyzed by normalizing to housekeeping gene beta-actin, and presented relative to in vitro cultured alveolar spheres.

Wu T., Rabi S.A., Michaud W.A., Becerra D., Gilpin S.E., Mino-Kenudson M, & Ott H.C. (2022). Protease inhibitor Camostat Mesyalte blocks wild type SARS-CoV-2 and D614G viral entry in human engineered miniature lungs. Biomaterials, 285, 121509.

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