Total RNA was extracted with miRneasy FFPE (QIAGEN, Venlo, Netherland)Kit followed by DNase I treatment.
Label
SYBR Green
Label protocol
PCR assays were performed using RT2 Profiler PCR Array (QIAGEN,Venlo, Netherland) , Rat Tumor Metastasis <PARN-028-ZD> , following the Manufacturer’s instructions. Reverse transcription was performed from 500 ng total RNA using the RT2 First Strand Kit (QIAGEN, Venlo, Netherland). Quantitative real-time PCR were performed (Light Cycler 480, Roche) 40 cycles of denaturation at 95˚C for 15 s; annealing and a final extension at 60˚C for 1 min.
Hybridization protocol
n/a
Scan protocol
n/a
Description
Test
Data processing
The normalization and all the data analysis were performed according to the manufacturers instructions using their web-based software package: http://sabiosciences.com/pcrarraydataanalysis.php For the normalization it uses the average of five housekeeping genes: ACTB, B2M, HPRT1, LDHA, RPL1 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.
