QIAseq miRNA Library Kit

The QIAseq miRNA Library Kit analysis workflow is based on miRBase, why only species present in miRBase are supported.

Ingenuity Pathway Analysis for miRNA data is restricted to human, mouse and rat. For this reason, biological insights results will be generated for these species only.

For an overview of what species are supported for the QIAseq miRNA Library Kit, see About analysis workflow versions and sample comparability.

Analysis workflow versions

Align and count analysis workflow parameters
Only applied options and parameters are listed.

Trim Reads (Thermo Fisher only)  
Adapter trimming by Trim adapter list 5prime_Ion_Torrent_miRNA_trim_adapter

Create UMI Reads for miRNA  
Minimum length of miRNA 15
Maximum length of miRNA 55
UMI length 12
Maximum size of small UMI groups 1
Common sequence AACTGTAGGCACCATCAAT
Maximum differences in common sequence 1

Quantify miRNA  
Minimum supporting count 1
Prioritized species Selected species
Allow length-based isomiRs Yes
- Additional upstream bases 2
- Additional downstream bases 2
- Missing upstream bases 2
- Missing downstream bases 2
Maximum mismatches 2
Strand specific Yes
Minimum sequence length 18
Maximum sequence length 25
Annotation records v1.0, v1.1: miRBase, piRNA, tRNA, rRNA, mRNA, snoRNA, and lncRNA/scRNA/ncRNA/snRNA
  v1.2, v2.0: miRBase, piRNA (when available)

RNA-Seq Analysis  
Mismatch cost 2
Insertion cost 3
Deletion cost 3
Length fraction 0.8
Similarity fraction 0.8
Auto-detect paired distances Yes
Maximum number of hits for a read 10
Strand specific Both
Library type Bulk
Ignore broken pairs Yes
Expression value Total counts

Create experiment analysis workflow parameters
Only applied options and parameters are listed.

Differential Expression for RNA-Seq  
Technology Small RNA
Normalization method TMM
Filter on average expression for FDR correction Yes

Create Heat Map for RNA-Seq  
Distance measure Euclidean distance
Linkage criteria Complete linkage