Total RNA was isolated from HSFs according to the manufacturer's instructions (Thermo Scientific). Single-stranded cDNA was synthesized using a kit from Thermo Scientific. Quantitative real-time PCR (qRT-PCR) was performed according to an miRNA protocol from Thermo Scientific using miR-217- or U6-specific primers (RiboBio Company, Guangzhou, China), inventoried real-time miRNA expression assays, and a RT-qPCR machine from Thermo Scientific. The comparative CT (ΔΔCT) method was used to calculate the real time quantitative (RQ) of miRNA expression, using U6 as a reference gene.
Rt qpcr machine
Manufactured by Thermo Fisher Scientific
Sourced in China, Denmark
The RT-qPCR machine is a laboratory instrument designed for quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) analysis. It is used to detect and quantify specific RNA molecules within a sample, enabling researchers to study gene expression and other related applications.
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Lab products found in correlation
2 protocols using rt qpcr machine
1
Quantitative Analysis of miRNA-217 Expression
2
Quantitative Real-Time PCR Protocol
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Total RNA was isolated at the indicated time points using RNeasy Plus Mini Kit (74134, Qiagen, Hilden, Germany). cDNA was prepared with 1 μg total RNA by High-Capacity cDNA Reverse Transcription Kit (4368814, Thermo Fisher Scientific). The reaction was diluted 20 times. Quantitative real-time PCR was performed according to manufacturer’s protocol (08-36-00001, TAG Copenhagen, Denmark) using an RTq-PCR machine (Thermo Fisher Scientific). Primers were designed using https://blast.ncbi.nlm.nih.gov/Blast.cgi accessed on 28 July 2018 and sequences can be obtained upon request.
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