GSM8183413: intestine_Trim28_light_normal_rep1; Mus musculus; RNA-Seq
Quick-DNA/RNA Microprep Plus Kit (Zymo) Libraries were prepared by the Van Andel Genomics Core from 142 ng of total RNA using the KAPA RNA HyperPrep Kit (Kapa Biosystems, Wilmington, MA USA). Ribosomal RNA material was reduced using the QIAseq FastSelect -rRNA HMR Kit (Qiagen, Germantown, MD, USA). RNA was sheared to 300-400 bp. Prior to PCR amplification, cDNA fragments were ligated to IDT for Illumina TruSeq UD Indexed adapters (Illumina Inc, San Diego CA, USA). Quality and quantity of the finished libraries were assessed using a combination of Agilent TapeStation 4200 (Agilent Technologies, Inc.) and QuantiFluor® dsDNA System (Promega Corp., Madison, WI, USA). Individually indexed libraries were pooled and 2x100 bp sequencing was performed on an Illumina NovaSeq6000 sequencer to an average depth of 50M raw paired-reads per transcriptome. Base calling was performed by Illumina RTA3 and output was demultiplexed and converted to FastQ format with Illumina Bcl2fastq2 v2.20.