IAB RNA-seq

The IAB RNA-seq product can be used for precise detection of differentially expressed genes (DEG)  from bacteria (using the NEBNext rRNA Depletion Kit, NewEngland BioLabs) to mammals (using the NEBNext poly(A) enrichment kit, New England BioLabs) with strand specificity. Thanks to our expertise with the preparation of RNA-seq libraries, IAB guarantees the preparation consistency over all samples, which is crucial for RNA-seq protocol.

IAB RNA-Seq workflow differs depending on if polyA mRNA enrichment or ribo-depletion is used. We follow the strict quality procedures to perform the essential QC steps on your samples and generate reliable results.

Sample requirement

• total RNA, gDNA free, no Mg 2+, EDTA/EGTA, guanidium, etc.
• RNA concentration: ≥ 10 ng/µl total RNA (polyA mRNA workflow), RIN≥7
•                                     ≥   5ng/ µl total RNA (rRNA depletion workflow), RIN≥7
• Minimum sample volume: 50 µl

Kit for library preparation

• NEBNext Ultra II Directional RNA Library Prep kit for Illumina (New England BioLabs)

Library QC and Pre-sequencing

• Library QC
• iSeq 100 (Illumina)
• Pair-end sequencing 2×100 bp
• Evaluation of equimolarity of libraries 
• Accurate determination of adapter-dimers
• Optimization of loading concentration for further sequencin

Sequencing

• • NovaSeq 6000 (Illumina)
  • Pair-end sequencing 2×100 bp
  • Internal standard – PhiX Spike-in
• BaseSpace Sequencing Hub for data storage and sharing 

Data QC

• Guaranteed ≥ 80% bases QC ≥ 30 (sequencing mode 2x100bp)

Bioinformatic analysis

• advanced data QC and statistics
• quantification of transcript abundance
• differential expression

Delivery time

• 8 weeks since sample delivery to the data output
• accelerated service on request wofkova@iabio.eu

Certification

• ČSN EN ISO 9001:2016
• ČSN EN ISO 13485 ed.2:2016