For transient depletion of DDX1, 30 pmol of short-interfering (siRNA) was electroporated into 3x106 CH12 cells using Amaxa Cell Line Nucleofector Kit R (Lonza). A pool of 4 siRNAs was used to target mouse DDX1 (SMARTpool: siGENOME DDX1 siRNA, Dharmacon) as well as a control siRNA ...
The reaction mixture contained 5 pmol of forward and reverse primer, 2× iQ SYBR Green Super Mix (Bio-Rad), and 2 μl of template cDNA. Standard curves were prepared for each gene using 5-fold dilutions of a known quantity (250 ng/μl) of genomic DNA from wild-type T. brucei DNA...