title: Illumina MiSeq paired end sequencing; COMPLEMENT C3aR ACTS AS AN ONCOSUPPRESSOR IN COLORECTAL CANCER BY MODULATING GUT MICROBIOTA
design: COMPLEMENT C3aR ACTS AS AN ONCOSUPPRESSOR IN COLORECTAL CANCER BY MODULATING GUT MICROBIOTA
library_name: S23_p
library_construction_protocol: C57BL/6J-ApcMin/J (referred to as APCMin/+) and C57BL/6J (referred to as WT) mice were bred and maintained in our SPF (Specific Pathogen Free) animal facility. APCMin/+/C3aR-/- mice were generated by crossing C57BL/6J-ApcMin/J with C3aR-/- mice in our animal facility. 12 week old APCMin/+, WT, C3aR-/- and APCMin/+/C3aR-/- mice (4mice/group) were sacrificed. The proximal (first 3 cm closer to the caecum) and distal colon (the final 3,5 cm closer to the rectum) and the single tumors were immediately preserved in 2ml of RNAlater stabilization solution (Qiagen) according to manufacturer instructions and stored at -20°C.  For RNA extraction, tumors and colons were homogenized in Trizol (500 ul/50 mg weight), centrifuged at 13000 rpm and the supernatant was loaded on Qiagen Mini Kit columns and treated as indicated in the manufacturer instructions. RNA concentration was quantified by Nanodrop and quality and integrity were evaluated by using a bioanalyzer (Agilent Technologies). Only samples with a RIN>8 were used for downstream applications. 1ug of RNA was used to prepare cDNA with TruSeq RNA kit by following the recommendations for low sample preparation protocol (Illumina)
attributes.tag: Experimental Factor: genotype
attributes.value: APCMin
attributes.tag: Experimental Factor: sampling site
attributes.value: distal colon