PBMC Total RNA Extraction and cDNA Synthesis

This procedure was performed as previously described (8 (link)). In brief, the total RNA from the PBMCs was extracted using a RNeasy mini kit (74104; Qiagen, Hilden, Germany) and quantified using a NanoDrop 3300 Fluorospectrometer (Thermo Fisher Scientific, Wilmington, DE, USA). cDNA was synthesized using a ReverTra Ace qPCR kit (FSQ-101; Toyobo, Kagoshima, Japan) and the reverse transcription (RT) conditions were as follows: 65°C for 5 min, 37°C for 15 min and 98°C for 5 min.

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PU D, & WANG W. (2014). Toll-like receptor 4 agonist, lipopolysaccharide, increases the expression levels of cytokines and chemokines in human peripheral blood mononuclear cells. Experimental and Therapeutic Medicine, 8(6), 1914-1918.

Publication 2014

RNeasy mini kit NanoDrop 3300 Fluorospectrometer ReverTra Ace qPCR kit

Cdna Reverse transcription

Corresponding Organization :

Other organizations : Sichuan University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Total RNA from PBMCs

control variables

Extraction of total RNA using RNeasy mini kit (74104; Qiagen, Hilden, Germany)
Quantification of total RNA using NanoDrop 3300 Fluorospectrometer (Thermo Fisher Scientific, Wilmington, DE, USA)
CDNA synthesis using ReverTra Ace qPCR kit (FSQ-101; Toyobo, Kagoshima, Japan)
Reverse transcription (RT) conditions: 65°C for 5 min, 37°C for 15 min and 98°C for 5 min

controls

No positive or negative controls explicitly mentioned

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