Bulk RNA Sequencing

Cloudbreak™ chemistry was used to perform bulk RNA sequencing (RNA-Seq) of human reference RNA samples with a control RNA spike-in. 

Methods 

The KAPA mRNA HyperPrep Kit prepared input of 100 ng Universal Human Reference RNA mixed with 1000x diluted External RNA Controls Consortium (ERCC) Mix 1 or Mix 2. Four replicates were prepared with ERCC Mix 1 and another four replicates were prepared with ERCC Mix 2. Sample messenger RNA (mRNA) was captured and fragmented before complementary DNA (cDNA) synthesis. The libraries were ligated with KAPA unique dual indexes (UDIs), amplified, and pooled equally. The AVITI™ System performed a 2 x 75 run and Bases2Fastq generated the FASTQ files.