cell type: PMA-differentiated THP-1 cell activation agent: Control time: 2 days
Extracted molecule
total RNA
Extraction protocol
Total RNA was extracted with Maxwell® RSC miRNA Tissue Kit (Promega)
Label
Taqman probes
Label protocol
Reverse transcription was performed from 500 ng total RNA using SuperScript VILO cDNA Synthesis Kit (Invitrogen). PCR were performed using TaqMan Fast Universal PCR Master Mix (2X), no AmpErase UNG (Applied Biosystem) following the Manufacturer’s instructions. Quantitative real-time PCR were performed with the QuantStudio 12K Flex (Applied Biosystems) with a 2 min hold at 50°C, a 10 min hold at 92°C, then 40 cycles of 3s 95°C denaturation followed by 30s of 60°C extention.
Hybridization protocol
n/a
Scan protocol
n/a
Description
Control
Data processing
The normalization was performed according to the manufacturers instructions and their web-hosted software https://apps.thermofisher.com/apps/spa/#/dashboard For the normalization it uses the average of five housekeeping genes: B2M, HPRT1 , RPL13A, GAPDH, ACTB. Target gene signals normalized to housekeeping genes; 2^-deltaCt, where deltaCt = (Ct_Target − Ct_HKG)] The web-based software package automatically performs all deltadeltaCt based fold-change calculations from the uploaded raw threshold cycle data.
