group1: test group2: test gender: male estimated days after hiv infection: 128
Extracted molecule
total RNA
Extraction protocol
Total RNA of PBMCs was isolated with Trizol reagent (Invitrogen). E coli polyA polymerase was used to add adenines at the 3' end of RNA molecules lacking a polyA tail. After oligodT annealing, a universal tag was attached to the 3' end of cDNAs during cDNA synthesis using retro-transcriptase Superscript III (Invitrogen).
Label
SYBR Green
Label protocol
With this universal tag, SYBR-based real-time quantitative PCR method was performed with miRNA-specific forward primers and a reverse universal primer mix.
Hybridization protocol
n/a
Scan protocol
n/a
Data processing
Preprocessing of raw data and statistical analyses were performed using RealTime StatMiner software. U1 and U6 were used in the training cohort and RNY3 and U6 was used in the validation cohort for normalization.
