Total RNA was extracted from PBMCs or cultured cells using M5 Hiper Universal Plus RNA Mini Kit (Mei5 Biotechnology, China). cDNA was synthesized using the cDNA synthesis kit or Mir-X miRNA First-Strand Synthesis Kit (both TaKaRa, Japan). Primers for qRT-PCR were designed via a public resource named PrimerBank (https://pga.mgh.harvard.edu/primerbank/ ) and synthesized by Sangon Biotech, which are listed in Table 3 . qRT-PCR amplification was performed using SYBR Green PCR Premix Ex Taq™ II reagents (TaKaRa, Japan) with the Quant Studio 6 FlexI real-time PCR system (Applied Biosystems, USA), following the protocols from the commercial kits. Expression levels of tested genes were determined with the 2-ΔCt or 2-ΔΔCt method based on the endogenous control (GAPDH or U6).
M5 hiper universal plus rna mini kit
Manufactured by Mei5 Biotechnology
Sourced in China
The M5 HiPer Universal Plus RNA Mini Kit is a laboratory equipment product designed for the efficient extraction and purification of high-quality RNA from a variety of sample types. It utilizes a silica-based membrane technology to capture and purify RNA molecules.
Automatically generated - may contain errors
4 protocols using m5 hiper universal plus rna mini kit
1
qRT-PCR Gene Expression Analysis
2
Quantitative Real-Time PCR Analysis
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Total RNA was extracted from cells by using the M5 HiPer Universal Plus RNA Mini Kit (Mei5 Biotechnology, China). cDNA was synthesized with the cDNA synthesis kit or Mir-X miRNA First-Strand Synthesis Kit (both were obtained from TaKaRa, Japan). Primers for qRT-PCR were designed (listed in Table S1
3
COPD Patient PBMC Isolation and RNA Extraction
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Peripheral blood samples from COPD patients and healthy controls were collected into EDTA-anticoagulated vacutainer tubes. PBMCs were isolated from 10 ml of blood sample per case by density gradient centrifugation using human lymphocyte separation medium (Solarbio Life Sciences, China) and were immediately stored at −80°C until the assay. Total RNA was extracted from the PBMCs of each case with the M5 HiPer Universal Plus RNA Mini Kit (Mei5 Biotechnology, China), according to the kit instructions. The concentration of total RNAs were measured with the Nanodrop ND-2000 spectrophotometer.
4
COPD Patient PBMC Isolation and RNA Extraction
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Peripheral blood samples from COPD patients and healthy controls were collected into EDTA-anticoagulated vacutainer tubes. PBMCs were isolated from 10 ml of blood sample per case by density gradient centrifugation using human lymphocyte separation medium (Solarbio Life Sciences, China) and were immediately stored at −80°C until the assay. Total RNA was extracted from the PBMCs of each case with the M5 HiPer Universal Plus RNA Mini Kit (Mei5 Biotechnology, China), according to the kit instructions. The concentration of total RNAs were measured with the Nanodrop ND-2000 spectrophotometer.
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