MicroRNA Sequencing
Cloudbreak™ chemistry performed microRNA (miRNA) sequencing of human reference RNA.
Methods
The QIAGEN QIAseq miRNA Library Kit prepared libraries from inputs of 100–200 ng. Six input replicates of Human Brain Reference (HBR) Total RNA (Thermo Fisher Scientific) and six replicates of Human XpressRef (XR) Universal Total RNA (QIAGEN) were prepared for 12 replicates total.
The Element Adept™ Library Compatibility Workflow adapted the replicates for sequencing on the Element AVITI™ System via two methods: the Adept Rapid PCR-Free protocol circularized the 12 replicates to generate circular libraries and the Adept Rapid PCR-plus protocol amplified the 12 replicates to generate linear libraries for onboard circularization. Libraries were pooled equally before sequencing, and AVITI performed a 1 x 72 run with two index reads consisting of 10 bp each. Bases2Fastq Software generated FASTQ files.
Reads were downsampled to 10 million per sample for analysis with the QIAGEN RNA-seq Analysis Portal. Downsampled and complete (no downsampling) data are available for each sample.