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RNA DISCOVERY SOLUTIONSProviding Sample to Insight

QIAGEN RNA DISCOVERY SOLUTIONS – PROVIDING SAMPLE TO INSIGHT

Reliable results in end-point RT-PCR, real-time RT-PCR, microarray and next-generation sequencing analysis depend strongly on the quality of the RNA sample used. To ensure success in your experiments, QIAGEN provides a comprehensive range of RNA sample technologies that deliver high-quality RNA from biological samples. Innovative advancements in the QuantiNova product range offering cutting-edge solutions for real-time PCR, minimizing experimental errors and artefacts. Array and NGS-based technologies allow researchers to gain more insight into individual samples using either a disease or pathway specific approach. Let us aid you in finding the best approach to generate relevant, actionable findings.

QIAGEN RNA WORKFLOW SOLUTIONS

RNA EXTRACTION AND PURIFICATION

END-POINT RT-PCR SOLUTIONS

TARGETED ARRAYS & PANELS

AUTOMATION SOLUTIONS FOR RNA ANALYSIS

SAMPLE COLLECTION AND STABILIZATION

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PRE-AMPLIFICATION FOR SINGLE CELL

ANALYSIS/LIMITED SAMPLE AMOUNTS

QUANTINOVA REAL-TIME PCR ASSAY KITS

GENEGLOBE PORTAL

01 | SAMPLE COLLECTION AND STABILIZATION

Once a biological sample is harvested, its RNA becomes extremely unstable. Immediate stabilization of cellular RNA is critical to minimize pre-analytical variables and to ensure accurate and reproducible analysis.

Sample Type

Kit

Human Blood

Tissue

Cells

Bacteria

Saliva

AnimalBlood

HumanBlood

HumanBoneMarrow

FixedTissue

• Allprotect Tissue Reagent• RNAlater Reagent• RNAlater TissueProtect Tubes• RNeasy Protect Mini and Midi Kits

• RNAprotect Cell Reagent• RNeasy Protect Cell Mini Kit

• RNAprotect Bacteria Reagent• RNeasy Protect Bacteria Mini Kit

• RNeasy Protect Saliva Mini Kit

• RNAprotect Animal Blood Tubes• RNeasy Protect Animal Blood Kit• miRNeasy Protect Animal Blood Kit

• PAXgene Blood RNA Tubes • PAXgene Blood RNA Kit• PAXgene Blood miRNA Kit

• PAXgene Bone Marrow RNA Tubes• PAXgene Bone Marrow RNA Kit

• PAXgene Tissue Containers• PAXgene Tissue RNA Kit• PAXgene Tissue miRNA Kit

02 | RNA EXTRACTION AND PURIFICATION

When purifying RNA it is essential to utilize a method that maintains RNA integrity and effectively removes gDNA, contaminants and inhibitors, all of which can affect downstream applications.

RNA incl. miRNA

Analyte RNA

Human Blood

Human Blood

Serum/plasma (cell-free body fluids*)

Tissue Easy to lyse

TissueDifficult to lyse

Cells or cell super-natants

Limited sample amount

FFPE Tissue Sections

Plant

AnimalBlood

Circulating nucleic acid

RNA clean-up

Exosomal RNA

RNA incl. gDNA

removal

DNA, RNA and Protein

Viral RNA

• QIAamp RNA Blood

• PAXgene Blood System

• RNeasy Plus

• RNeasy Plus Universal• RNeasy Lipid Tissue• RNeasy Fibrous Tissue

• RNeasy Plus

• RNeasy Micro

• RNeasy FFPE

• RNeasy Plant

• RNeasy MinElute

• PAXgene Blood System

• RNeasy MinElute

• RNeasy MinElute

• miRNeasy Serum/Plasma

• exoRNeasy

• QIAamp Viral RNA Mini• QIAamp DSP Virus• QIAamp MinElute Virus Spin• QIAamp MinElute Virus Vacuum• QIAamp UltraSens Virus

• QIAamp Circulating Nucleic Acid

• RNeasy Plus Universal

• miRNeasy • RNeasy Plus

• miRNeasy

• miRNeasy • exoRNeasy • RNeasy Plus

• AllPrep DNA/RNA Micro • Allprep DNA/RNA Mini • AllPrep RNA/Protein • AllPrep DNA/ RNA/Protein Mini

• miRNeasy Micro

• RNeasy Plus Micro

• miRNeasy FFPE • RNeasy FFPE

• miRNeasy

• RNeasy Protect Animal Blood

• RNeasy Protect Animal Blood

• RNeasy Protect Animal Blood

• RNeasy Protect Animal Blood

• RNeasy MinElute

• RNeasy MinElute

• RNeasy MinElute

• RNeasy MinElute

• RNeasy MinElute

• RNeasy MinElute

03 | PRE-AMPLIFICATION FOR SINGLE CELL ANALYSIS/LIMITED SAMPLE AMOUNTS

Pre-amplification overcomes the limitations imposed by limited RNA availability, enabling downstream investigation of the transcriptome with minimal bias.

REPLI-g WTA Single Cell Kit For whole transcriptome amplification of total RNA or mRNA from single cells (1-1000 cells).

REPLI-g Single Cell RNA Library Kit (Illumina) Contains novel REPLI-g SensiPhi DNA Polymerase, as well as an optimized set of buffers and reagents for whole transcriptome amplification (WTA) from just single cells (up to 1000 cells) or equivalently small samples.

REPLI-g Cell WGA & WTA Kit For parallel whole genome and whole transcriptome amplification from cells and limited samples (25-1000 cells).

RT² PreAMP cDNA Synthesis Kit For pre-amplification of cDNA from small or FFPE samples of RNA for analysis on RT² Profiler PCR Arrays.

miScript PreAMP PCR Kit For pre-amplification of cDNA from samples containing low RNA amounts prior to real-time PCR analysis of miRNA using miScript PCR Arrays.

04 | END-POINT RT-PCR SOLUTIONS

QIAGEN’s new OneStep Ahead RT-PCR Kit offers faster one-step RT-PCR (1-hour protocol) with high sensitivity, specificity and fidelity. The unique two-phase hot-start procedure allows for room-temperature setup and optional pipetting control offer increased convenience. The dedicated enzyme mix improves specificity, sensitivity (0.1pg to 2µg) and fidelity.

05 | QUANTINOVA REAL-TIME PCR ASSAY KITS

QIAGEN’s new range of QuantiNova kits offer a breakthrough in real-time PCR technologies. In-process safety features assist in eliminating variables and minimizing artefacts. QuantiNova kits offer the added convenience of room temperature setup and in-built visual pipetting controls. QuantiNova kits are compatible will all real-time PCR instruments, irrespective of format, and provide consistent, sensitive and reproducible results.

QuantiNova Reverse Transcription KitAllows for fast cDNA synthesis and efficient removal of contaminating gDNA through the use of an integrated gDNA removal mix. Includes internal control RNA to assess for successful reverse transcription and amplification (requires separate QuantiNova IC assay and appropriate assay kit for detection).

QuantiNova SYBR Green PCR Kit Accurate and robust detection down to single target copy targets using ultrafast cycling protocol. Superior sensitivity and reaction efficiencies.

QuantiNova Probe PCR Kit Highly sensitive, specific, and ultrafast, probe-based real-time PCR. Duplex capability. Robust detection down to single copy targets. Stable for up to 100 hours at 30�C.

QuantiNova SYBR Green RT-PCR KitUnique two-phase hot-start procedure for room temperature setup. Internal control for positive in-process verification of successful RT-PCR. Ultrafast, universal protocol for all standard and fast cyclers.

QuantiNova Probe RT-PCR Kit One-step qRT-PCR using sequence-specific probes for gene expression analysis. Unique two-phase hot-start procedure for room temperature setup. gDNA reduction for improved quantification accuracy.

QuantiNova IC Probe Assay (primer/probe mix) Detects QuantiNova Internal Control RNA; use with QuantiNova Probe PCR Kit or QuantiNova Probe RT-PCR Kit.

QuantiNova IC SYBR Green Assay (primer mix) For SYBR-based detection of QuantiNova Internal Control RNA; use with QuantiNova SYBR Green PCR Kit or QuantiNova SYBR Green RT-PCR Kit.

QuantiTect Primer AssaysGenome-wide, bioinformatically validated primer sets for use in SYBR Green-based real-time RT-PCR on any cycler. Assays are available for all genes from human, rat, mouse, and many other species.

Isolate RNA from research samples:cells, tissues, FFPE, and/or blood

Add cDNA to RT²SYBR Green qPCR Mastermix; aliquot accross RT²Profiler PCR

Array

Convert totalRNA to cDNA with RT² First Strand Kit

Generate data in anyreal-time PCR instrument

Analyse your data

Interpret your data and publish your results

Control Experimental

06 | TARGETED ARRAYS & PANELS

Arrays allow researchers to evaluate more data points in a single experiment, reducing sample input requirements. • RT² Profiler PCR Arrays • miScript PCR System • QIAseq Targeted RNA and Targeted Virtual Panels

06.01 | RT² PROFILER PCR ARRAYS

Data analysis is simplified with an easy to use Microsoft Excel analysis macro. Alternatively, researchers can utilize the GeneGlobe Data Analysis Center: • Intuitive, user-friendly interface with no bioinformatics skills needed • Complimentary data analysis for all GeneGlobe assays and arrays • Get results in scatter plot, volcano plot, clustergram and heat map formats

RT² Profiler PCR Array Plate Layout

Housekeepinggenes

GenomicDNA

control

Reversetranscription

controls

PositivePCR controls

PCR Arrays are the most reliable tools for analyzing the expression of a focused panel of genes. Each 96-well plate, 384-well plate, or 100-well disc PCR Array includes SYBR Green-optimized primer assays for a thoroughly researched panel of relevant, pathway- or disease-focused genes. PCR Arrays can also be customized to contain a panel of genes tailored to your specific research interests. The simplicity of the PCR Arrays makes them accessible for routine use in every research laboratory.

RT² qPCR Primer Assays are designed, bench validated, and quality controlled to ensure sensitive and specific amplification. Controls are also included on each array for genomic DNA contamination, RNA quality, and general PCR performance. Individual assays are available separately.

06.02 | MISCRIPT PCR SYSTEM

miScript miRNA PCR Arrays contain miScript Primer Assays that have been arrayed into full miRNome, high content, and biologically or disease relevant panels. All miScript Arrays are partially or fully customisable for content.

06.03 | QIASEQ TARGETED RNA AND TARGETED VIRTUAL PANELS

QIAseq Targeted RNA Panels have been developed as a “Sample to Insight” solution for quantitative gene expression profiling using RNAseq. These panels integrate molecular barcode technology and a two-stage PCR-based library preparation to deliver unbiased and accurate quantification for your digital RNA sequencing results. These panels can be used with any Illumina or Thermo-Fisher NGS instrument.

The QIAseq Targeted Panels are wet-bench verified to ensure the highest quality of results.

The QIAseq Targeted RNA Virtual Panels build on QIAGEN’s know-how to deliver more than 170 panels with disease- and biologically-relevant gene content. These panels have been curated over several years by mining literature and accessing QIAGEN's proprietary knowledge database. Researchers can add up to 25 genes to a catalogued panel.

Disadvantages QIAseq Targeted RNA Panels

Human Blood

Whole transcriptome sequencing (WTS)

Microarrays

qPCR and ddPCR

TraditionaltargetedRNAseq

• Expensive• Complex data analysis

• Cost effective• Simplified data analysis• Compatible with FFPE samples

• High background noise• Low dynamic range• High RNA requirement

• High specificity• High dynamic range• Low RNA requirement

• Limited sample and assay throughput• Requires lots of RNA

• High sample and assay throughput• Requires as little as 25ng RNA

• Inaccurate gene expression profiling due to PCR duplicates and amplification bias

• Digital quantification enabled by molecular barcodes for precise gene expression profiling

07 | GENEGLOBE PORTAL

At GeneGlobe, you can discover and order gene- and pathway-specific solutions for qPCR, mRNA, lncRNA, miRNA gene expression analysis, next-generation sequencing, microbial detection, RNAi, genotyping, epigenetics and functional analysis. Browse the product range according to your gene, pathway, technology or biological field of interest.Visit www.qiagen.com/za/geneglobe

GENEGLOBE PRODUCT

PORTFOLIO

NGS

siRNA

CellSignaling

GenotypingMicrobial

Epigenetics

miRNA

GeneExpression

Targeted sequencing• QIAseq DNA panels• QIAseq RNA panels*• QIAseq miRNA panels*• QIAseq fusion gene panels*

siRNA• FlexiTube® siRNA• FlexiPlate• shRNA plasmids

Cell-based reporters• Cignal™ reporter arrays/assays

Copy number and Somatic mutation• qBiomarker® Copy Number arrays/assays• qBiomarker Somatic Mutation arrays/assays

Microbiome• qPCR arrays/assays• qPCR multi-assays/kits

DNA methylation• Pyromark® CpG assays• EpiTect® Methyl II arrays/assays• EpiTect ChiP arrays/assays*

miRNA gene expression• Expression arrays/assays• Mimics and inhibitors• Target protectors

mRNA gene expression• RT² PCR arrays/assays• QuantiTect® Primer assayslncRNA gene expression• RT² lncRNA PCR arrays/assays

* Coming soon

08 | AUTOMATION SOLUTIONS FOR RNA ANALYSIS

QIAcubeAutomation of QIAGEN spin-column processing (12-sample throughput).

QIAcube HT Enables automated mid- to high-throughput nucleic acid purification in 96-well format using silica membrane technology.

QIAsymphony SP Automated nucleic acid extraction, combines the speed and efficiency of silica-based purification with the convenient handling of magnetic particles (96-sample throughput).

QIAgility Compact benchtop instrument that enables rapid, high-precision setup of PCR experiments with ready-to-use software for ease and convenience.

QIAxpert Innovative high-speed microfluidic UV/VIS spectrophotometer which profiles sample content to differentiate between DNA, RNA and sample impurities.

QIAxcel Advanced Fully automates sensitive, high-resolution capillary electrophoresis of up to 96 samples per run. The QIAGEN RNA Integrity Score (RIS) provides an objective quality measurement of the analyzed samples and allows easy interpretation of sample integrity.

Rotor-Gene QOffers outstanding thermal and optical performance for real-time PCR. Unmatched optical range. Supports all state of the art analyses.

Automatedsolutions for RNA

Analysis

Human Blood

BiologicalSampleDisruption

SamplePurification

QualityControl

AssaySetup

• TissueLyser• TissueLyser II• TissueRuptor

• QIAcube• QIAcube HT• QIAsymphony SP

• QIAxpert• QIAxcel Advanced

• QIAgility

Detection &Analysis

• Rotor-Gene Q• QIAxcel Advanced

Proud supplier of QIAGEN in South Africa:

Whitehead Scientific (Pty) LtdSouth Africa - Cape Town(tel) +27 (0) 21 944 6460 - (fax) +27 (0) 21 949 5478PO Box 194, Brackenfell, 7561, Cape Town, South Africawhitesci@whitesci.co.za - www.whitesci.co.za

RNA Workflow Solutions Brochure 04/2016