RNA was isolated using TRI reagent solution (Sigma) followed by the on‐column RNeasy mini kit and DNase treatment (Qiagen). cDNA synthesis was performed using the Transcription First Strand cDNA Synthesis Kit (Roche). Quantitative real‐time polymerase chain reaction (qPCR) was performed using the ABI 7900T PCR System (Applied Biosystems). The gene expression was normalized to the expression of housekeeping gene 18S. Relative fold changes were transformed using the comparative threshold cycle (CT) method (the 2‐ΔΔCT method) with the Sequence Detection System V2.4 software (Applied Biosystems). Primer sequences are provided in Supporting Information Table S1 .
Abi 7900t pcr system
Manufactured by Thermo Fisher Scientific
Sourced in United States
The ABI 7900T PCR System is a real-time polymerase chain reaction (PCR) instrument designed for quantitative gene expression analysis. It features a high-density thermal block and supports a wide range of sample volumes and formats. The system is capable of performing a variety of PCR applications, including gene expression profiling, microRNA analysis, and SNP genotyping.
Automatically generated - may contain errors
Lab products found in correlation
Rneasy mini kit, by Qiagen (3 mentions)
Tri reagent solution, by Merck Group (3 mentions)
Transcription first strand cdna synthesis kit, by Roche (2 mentions)
Dnase treatment, by Qiagen (2 mentions)
Sequence detection system v2, by Thermo Fisher Scientific (1 mentions)
Transcriptor first strand cdna synthesis kit, by Roche (1 mentions)
Mircury rna isolation kits biofluids, by Qiagen (1 mentions)
3 protocols using abi 7900t pcr system
1
Gene Expression Analysis by qPCR
2
RNA Isolation and qRT-PCR Analysis
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RNA was isolated using TRI reagent solution (Sigma) followed by the on-column RNeasy mini kit and DNase treatment (Qiagen, Germany). cDNA synthesis was performed using the Transcription First Strand cDNA Synthesis Kit (Roche). qRT-PCR was performed using ABI 7900T PCR System (Applied Biosystems). Gene expression using SYBR Magic was normalized to the expression of β-actin. The primers used in the present study were supplied in Supplementary Table S4 .
3
Extracellular Vesicle RNA Profiling
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RNA was extracted using TRI reagent solution (Sigma) and RNeasy mini kit (Qiagen, Germany). RNA was also purified with miRCURY RNA Isolation Kits-Biofluids (Exiqon) from re-suspended EVs. Complementary DNA (cDNA) synthesis was performed using Transcriptor First Strand cDNA Synthesis Kit (Roche for mRNA and Novabio for miRNA). qPCR assay was performed using ABI 7900 T PCR System (Applied Biosystems, Foster City, USA). The sequences of the primers used for detection are provided in Additional file 1 : Table S1.
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