Superscript 3 fs synthesis supermix kit

DNA and RNA were extracted from CLL PBMC samples using a standard kit according to the manufacturer’s protocol (Qiagen, Hilden, Germany). cDNA synthesis was performed using the Superscript III FS synthesis supermix kit (Life technologies, Carlsbad, USA) according to the manufacturer’s protocol. Real-time PCR was performed using the Power SYBR Green PCR master mix and the 7900HT fast real-time PCR instrument (Applied Biosystems, Warrington, UK). qRT-PCR primers were designed with PRIMER3 (Broad Institute). The GAPDH gene was used as an internal loading control gene. The relative expression levels were calculated using the ΔΔCt method. All primers sequences are listed in Supplemental Table 2.