Genomics & Analytical Services | Next Generation Sequencing

RNA-Seq TechnicalSpecifications
Azenta Life Sciences RNA Sequencing Services• Standard RNA-Seq• Single-Cell RNA-Seq*• Strand-Specific RNA-Seq• Iso-Seq*• Small RNA-Seq• Digital Spatial Profiling*• Ultra-Low Input RNA-Seq*Not covered here. See genewiz.com for more details.

RNA Sequencing Workflow
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Experimental
DesignSamplePreparation

Library
Preparation

Sequencing

Data Analysis

Project Delivery

Define objectives
and design assay
accordinglyExtract and purify
input RNA

Enrichment, cDNA
synthesis, andadapter ligation

Sequence libraries
using NGS
platforms

Trim, filter, andmap reads; perform
advanced analysis

Receive data files
and post-delivery
support112

Experimental Design

Azenta Life Sciences provides resources to help you
find the best NGS solution and experimental designfor your project.

56Sample Preparation

Sample Type*

Minimum Amount†

Recommended
AmountTotal RNA‡

500 ng (standard)
10 pg (ultra-low)

2 μgInteractive NGS Solution Selection

104 cells (standard)Tool: genewiz.com/ngs

Eukaryotic cell pellet

Contact us for a free technical

Prokaryotic cell pellet

106 cells

108 cells

Frozen tissue

2 mg

10 mg

FFPE

2 slides4 slidesconsultation with a Ph.D.-levelscientist

1 cell (ultra-low)

106 cells

Other sample types accepted. View Sample Submission Guidelines for details.
Please inquire about submitting lower inputs.
‡Contact us about Azenta Life Sciences’ RNA Stabilization Tubes to ship RNA samples atambient temperature.
*†azenta.com

Genomics & Analytical Services | Next Generation Sequencing

RNA-Seq Technical
Specifications
3

Library Preparation

RNA-Seq ServiceTarget RNA

RNA Selection Method

mRNA (eukaryotic)

Poly(A) selection

mRNA + lncRNA

rRNA depletion

Small

Small RNA (miRNA, siRNA, piRNA)

Size fractionation with adapter ligation to5’ phosphate

Ultra-Low Input

mRNA (eukaryotic)

Poly(A) selection with enrichment for
full-length transcripts

Standard & Strand-Specific

45Sequencing

Platform

Illumina® NovaSeq™ or HiSeq®

Configuration

2x150 bp

RNA-Seq Service

Depth

Customizable to your project needs*

Data Quality

Guaranteed ≥80% bases with Q30 or higher

Standard
Strand-SpecificUltra-Low Input*Generally, we recommend 5-10 million read pairs per sample for small genomes(e.g. bacteria) and 20-30 million read pairs per sample for large genomes (e.g.
human, mouse). Medium genomes often depend on the project, but 15-20 million
read pairs per sample is typically sufficient. For de novo transcriptome assembly
projects, we recommend 100 million read pairs per sample.

Project Delivery

Standard Analysis Additional Analysis
PackageOptions• RNA SNP/INDEL detection• Mapping• Differential geneexpression• Novel transcript discovery• De novo transcriptome
assembly• Mapping
Small•D ifferential gene
expression• Small RNA

Deliverables for All Projects

Data Delivery Options• Gene fusion discovery• Trimming• Trimming

6Data Analysis

Optional Deliverables• Aligned data (BAM file)• Hit counts (TXT file)

SFT P• Sample quality control report

SFTPCustomer
CloudAccountExternal
Hard Drive(US Only)• Raw data (FASTQ files)• DGE results (CSV file)• GO enrichment analysis (CSV file)• Differential splicing analysis(DEXSeq report)• De-multiplexed, aggregated Picard
BAM file with summary metrics

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