Illumina HiSeq 2500 sequencing; Elucidation of FOXP1 target genes in DLBCL by RNA-Seq
Elucidation of FOXP1 target genes in DLBCL by RNA-Seq
The DLBCL cell lines used were SUDHL6 of GCB DLBCL subtype and U-2932 of ABC DLBCL. Cell lines were maintained at 37 degrees C, 5%CO2 in a humidified atmosphere in RPMI supplemented with 20% heat-inactivated FBS and antibiotics.  The quality of the isolated RNA was determined with a Qubit® (1.0) Fluorometer (Life Technologies, California, USA) and a Bioanalyzer 2100 (Agilent, Waldbronn, Germany). Only those samples with a 260 nm/280 nm ratio between 1.8-2.1 and a 28S/18S ratio within 1.5–2 were further processed. The TruSeq RNA Sample Prep Kit v2 (Illumina, Inc, California, USA) was used in the succeeding steps. Briefly, total RNA samples (100-1000 ng) were poly A enriched and then reverse-transcribed into double-stranded cDNA. The cDNA samples was fragmented, end-repaired and polyadenylated before ligation of TruSeq adapters containing the index for multiplexing Fragments containing TruSeq adapters on both ends were selectively enriched with PCR. The quality and quantity of the enriched libraries were validated using Qubit® (1.0) Fluorometer and the Caliper GX LabChip® GX (Caliper Life Sciences, Inc., USA). The product is a smear with an average fragment size of approximately 260 bp. The libraries were normalized to 10nM in Tris-Cl 10 mM, pH8.5 with 0.1% Tween 20.
Experimental Factor: U-2932
cell line
Experimental Factor: FOXP1 siRNA 2
RNA interference