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Qs5 384 well pcr system

Manufactured by Thermo Fisher Scientific
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The QS5 384-well PCR system is a compact, high-throughput real-time PCR instrument designed for gene expression analysis, genotyping, and other PCR-based applications. It features a 384-well plate format and can perform up to 4 experiments simultaneously. The system provides accurate temperature control and optical detection for reliable real-time PCR results.

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Lab products found in correlation

Miscript 2 rt kit, by Qiagen (2 mentions) Power sybr green master mix, by Thermo Fisher Scientific (1 mentions) Qscript cdna synthesis kit, by Quantabio (1 mentions) Quick rna miniprep kit, by Zymo Research (1 mentions) Qscript kit, by Quantabio (1 mentions)

2 protocols using qs5 384 well pcr system

1

Quantitative Analysis of miRNA Expression

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Total RNA was extracted using the Zymo Quick-RNA MiniPrep Kit according to the manufacturer’s protocol. For pri- and pre- miRNAs and mRNAs, RT was performed using 200 ng of RNA with a Qscript cDNA Synthesis Kit (QuantaBio). For mature miRNAs, the RT reaction was performed using 200 ng of RNA with the miScript II RT Kit in a total volume of 20 μl (Qiagen). Subsequent qPCR analysis (see Supplementary Table 2 for primer sequences and Supplementary Figs. 2–7 for primer validation) using Power SYBR Green Master Mix (Life Technologies) or a TaqMan assay and a Applied Biosystems QS5 384-well PCR system (software v.1.3.0). For mature miR-155 levels, TaqMan assays were performed using the ipu-miR-155 Taqman Assay (Thermo Fisher Scientific, 467534). Data were analysed using the ΔΔCt method as described previously13 (link).
Tong Y., Lee Y., Liu X., Childs-Disney J.L., Suresh B.M., Benhamou R.I., Yang C., Li W., Costales M.G., Haniff H.S., Sievers S., Abegg D., Wegner T., Paulisch T.O., Lekah E., Grefe M., Crynen G., Van Meter M., Wang T., Gibaut Q.M., Cleveland J.L., Adibekian A., Glorius F., Waldmann H, & Disney M.D. (2023). Programming inactive RNA-binding small molecules into bioactive degraders. Nature, 618(7963), 169-179.
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2

Dose-dependent miRNA-200c regulation

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After transfection, cells were allowed to recover for 8 h in complete growth medium before splitting into 12-well plates (100,000 cells/well). After adhering for 12 h, the cells were treated with 1 or TGP-200c in dose response, vehicle (DMSO), LNA-200c, or LNA-Scramble for 60 h. Total RNA was extracted using the Zymo Quick-RNA Miniprep Kit per the manufacturer’s protocol, including DNase treatment. Reverse transcription (RT) for mature miRNAs was completed with 300 ng of total RNA using a miScript II RT Kit (Qiagen) per the manufacturer’s protocol. RT of pre-miR-200c and mRNAs was completed using qScript Kit (Quanta Bio) with 1000 ng of total RNA per the manufacturer’s protocol. qPCR was completed using a QS5 384-well PCR system (Applied Biosystems) using Power SYBR Green Master Mix according to the manufacturer’s protocol and the Comparative Ct with melt standard method, adjusted for a sample volume of 10 μL. All data were analyzed using the ΔΔCt method previously described (Rao et al., 2013 (link)).
Haniff H.S., Liu X., Tong Y., Meyer S.M., Knerr L., Lemurell M., Abegg D., Aikawa H., Adibekian A, & Disney M.D. (2021). A structure-specific small molecule inhibits a miRNA-200 family member precursor and reverses a type 2 diabetes phenotype. Cell chemical biology, 29(2), 300-311.e10.
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