The TRIzol reagent (Invitrogen, USA) was used for the extraction of total RNA from cells and tissues. A gDNA kit (TransGen Biotech, Beijing, China) was used to isolate gDNA from cells. The reverse transcription reaction utilized 1st Strand cDNA Synthesis SuperMix (gDNA Digester Plus) (Yeasen). For the RT-qPCR, 2 μL of cDNA template was employed with qTOWER3 (link) system (Analytik Jena, Germany) using qPCR SYBR Green Master Mix (Yeasen). GAPDH and U6 were used as internal controls to standardize the transcript levels based on the 2-△△Ct method. Primers employed in this study were synthesized by Sangon Biotech from Shanghai, China (Shanghai, China) (Table S1 ).
Qtower3 link system
Manufactured by Analytik Jena
Sourced in Germany
The QTOWER3 is an automated system for the extraction and purification of nucleic acids (DNA and RNA) from a variety of sample types. The system utilizes magnetic bead-based technology to perform all necessary steps, including sample lysis, binding, washing, and elution, in a fully automated manner. The QTOWER3 is designed to offer high throughput and reproducible results for a wide range of applications, such as diagnostic testing, research, and forensics.
Automatically generated - may contain errors
Lab products found in correlation
Nanodrop 1000, by Thermo Fisher Scientific (2 mentions)
Trizol, by Thermo Fisher Scientific (2 mentions)
Gdna digester plus, by Yeasen (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Qpcr sybr green master mix, by Yeasen (1 mentions)
Primescript rt reagent kit, by Takara Bio (1 mentions)
2 protocols using qtower3 link system
1
Transcriptome Analysis via RT-qPCR
2
Quantitative Gene Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Briefly, total RNA was extracted by TRIZOL (Thermo Fisher) following the manufacture’s instruction and quantified by Nanodrop 1000 (Thermo Fisher). The reverse transcription was carried out using the PrimeScript™RT reagent Kit (TAKARA, RR047Q) following the manufacture’s instruction. Quantitative PCR was carried out in qTOWER3 (link) system (Analytikjena) using TAKARA TB Green II Real-Time PCR Master Mix following the manufacture’s instruction. Quantitative PCR was performed with indicated primer for specific genes, and GAPDH served as control. The relative expression level was determined by −ΔΔCt method. qPCR primers are listed in Supplementary Data 7 .
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