Understanding Gene Expression and Epigenetics Using PCR Arrays

Understanding Gene Expression and
Epigenetics Using a Pathway Approach:
PCR Array Systems for Gene & MicroRNA
Expression Profiling
CpG Island Methylation Analysis
and ChiP

Samuel J. Rulli Jr., Ph.D.
qPCR Applications Scientist
Samuel.Rulli@QIAGEN.com
The PCR Arrays are intended for molecular biology applications. This product is not Intended for the
diagnosis, prevention, or treatment of disease.
Profiling genes by pathways and diseases

-1-

Sample & Assay Technologies

Understanding Gene Expression and
Epigenetics Using a Pathway Approach:
PCR Array Systems for Gene & MicroRNA
Expression Profiling
CpG Island Methylation Analysis
and ChiP

Questions, Comments, Concerns?
US Applications Support
support@sabiosciences.com

Global Applications Support
SABio@qiagen.com


-2-


Seminar Overview

Cancer Research

Apoptosis

What is Epigenetics?
PCR Arrays for Gene Expression
Profiling Tumor and Metastatic Tumor Cells

Cell Cycle

Inflammation

ECM/Adhesion

Neuroscience

Signal Transduction

PCR Arrays for Epigenetic Applications
Profiling miRNA Biomarkers in Colon Cancer
Profiling DNA Methylation in Breast Cancer Cell
lines and Normal Breast Tissue
Profiling Chromatin Structure through Histone
Modifications
Summary and Questions

Stem Cell/
Development
Toxicology/
Drug ADME


-3-


Seminar Overview

Cancer Research

Apoptosis


Cell Cycle

Inflammation

ECM/Adhesion

Neuroscience

Signal Transduction

Modifications

Stem Cell/
Development
Toxicology/
Drug ADME

The applications presented here are for research purposes.
Not for diagnostic use.

-4-



Epigenetics:
“Epi” from Greek meaning “above or over genetics”

Biological Definition:
Something that effects the Phenotype without
changing the Genotype;
Something that can effect gene expression without
causing a mutation in the DNA code


-5-


Current View of Gene Expression
Activated
Transcription Factors

RNAi:
miRNA

Protein “A”
NFκB

+

p53

Transcription
Initiation Complex

mRNA ”A”

–
Histones

Histones

Structural Gene “A”
Me

p53 BS

AC AC

Me

NFκB BS

Me Me

Me Me Me

Me Me

TSS (+1)

Gene Expression is Complex with multiple layers of control

-6-


Multiple Epigenetic mechanisms influence
Gene Expression
Activated

RNAi:
miRNA

NFκB
p53

Transcription
Initiation Complex

mRNA ”A”

Histones

Histones

Me

p53 BS

AC AC

Me

NFκB BS

Me Me

Me Me Me

Me Me

TSS (+1)


-7-


PCR Arrays for Gene Expression and Epigenetic
Regulation
Activated

RNAi:
miRNA

Protein “A”
NFκB
p53

Transcription
Initiation Complex

mRNA ”A”

Histones

Histones

Me

p53 BS

AC AC

Me

NFκB BS

Me Me

Me Me Me

Me Me

TSS (+1)


-8-


Epigenetics effects multiple pathways and diseases
Biological Mechanisms of Epigenetics:
miRNA
Regulates translation or stability of mRNA
DNA Methylation:
Regulates transcription factor access, chromatin structure
Histone modifications:
Regulates bulk chromatin structure and transcription factor access
Biological Pathways and Diseases Effected by Epigenetics:
Cancer- methylation of tumor suppressor genes
Differentiation- changes in the histone code
Immunology: miRNA regulation of cell cycle proteins by viruses
Almost all pathways and diseases may have an underlying Epigenetic component

HOW CAN RESEARCHERS PROFILE GENE EXPRESSION AND
EPIGENETIC MECHANISMS?

-9-


ANSWER: PCR Arrays
WHAT IS A PCR ARRAY?
Collection of SYBR-Green based qPCR Assays with biological content with
Focus on a Disease or Pathway
Available for any qPCR instrument
Available for a variety of laboratory applications:
mRNA profiling (Gene Expression)
miRNA profiling
DNA Methylation
Chromatin Precipitation
Part of a complete “System”
Sample Isolation to Final Data Publishable Data Component


- 10 -


5 Components in the PCR Array System

PCR ARRAYS are a Complete System for
Pathway-Focused Gene Expression Analysis
1.

Sample Isolation (QIAGEN RNeasy)*

2.

RT2 PCR Arrays (96, 384 and Rotorgene Q formats)

3.

RT2 First Strand kit**

4.

RT2 SYBR Green Master Mix

5.

Free data analysis software (web or excel-based)

Publishable story
Ease of use
Relevant genes
Accurate
Reliable

Complete System from Sample Preparation to Fold Change Results
*substitute QIAGEN miRNeasy ** substitute miScriptII RT Kit for miRNA
*substitute QIAGEN DNeasy Blood and Tissue; **substitute EpiTect Methyl II RE Kit


- 11 -


Seminar Overview

Cancer Research

Apoptosis


Cell Cycle

Inflammation

ECM/Adhesion

Neuroscience

Signal Transduction

Modifications

Stem Cell/
Development
Toxicology/
Drug ADME


- 12 -



Cancer Research

Research Question:

What are the gene expression differences between a
metastatic and non-metastatic Breast Tumor?


- 13 -


Anatomy of a RT2 Profiler PCR Array (for mRNA Profiling)

84 Pathway-Specific Genes of
Interest

.

5 Housekeeping Genes

.

Genomic DNA Contamination
Control
.

Reverse Transcription Controls
(RTC)
.

Positive PCR Controls (PPC)

.


- 14 -


Anatomy of a RT2 Profiler PCR Array (for mRNA Profiling)

84 Pathway-Specific Genes of
Interest
.

5 Housekeeping Genes

.

Genomic DNA Contamination
Control
.

Reverse Transcription Controls
(RTC)
.

Positive PCR Controls
(PPC)

.

.


- 15 -


Built-in Standard and Performance Controls
5 Housekeeping Genes Controls
Flexibility - researchers select the most stable reference (“housekeeping”) gene for their
experimental situation
Human: GAPDH, ACTB, B2M (Beta-2-Microglobulin), HPRT1 (Hypoxanthine
Phosphoribosyltransferase 1), RPL13A (Ribosomal Protein 13A)
Sample and qPCR Performance Controls
Reverse Transcription Controls (RTC)
Control for impurities that may degrade RNA or inhibit RT enzyme
Positive PCR Controls (PPC)
Control for inhibitors of the PCR step
Used in conjunction with RTC assay to establish RT efficiency
Genomic DNA Contamination Control (GDC)
Detect gDNA contamination
– Single exon genes important for biology (can’t design cross-intron qPCR
Assays)
– Pseudo-genes confound analysis of low expressing genes (detected with
cross-intron assays)
– Incorporated DNA removal step in cDNA synthesis kit to remove residual
genomic DNA

- 16 -


How RT2 Profiler PCR Arrays Work
cDNA Synthesis (C-03 kit)
– Genomic DNA Removal Step (5 min.)
– Reverse Transcription Step (20 min.)

.

Load Plates
1 Sample per PCR Array
2 minutes with multi-channel pipet

.

Run 40 cycle qPCR Program
Standard cycling conditions for all
Real Time PCR Instruments
2 hours

.

Upload and Analyze Data
15 minutes from Raw Ct to
Fold Change Data

.


- 17 -



.

Load Plates

.

2 hours

.

Fold Change Data

.


- 18 -



.

Load Plates

.

2 hours

.

Fold Change Data

.


- 19 -



.

Load Plates

.

2 hours

.

Fold Change Data

.


- 20 -


Clustergram
Data

Analysis

Web-Based Software
Scatter Plot
No installation or licensing needed

.

From Raw Ct Values to Fold Change Results
∆∆C(t) Method

.

Multiple Analysis Formats
Scatter Plot
Volcano Plot
Multi-Group Plot
Clustergram

.

Volcano Plot
Also available as downloadable Excel templates


- 21 -


Wet Bench QC Validation of ALL qPCR Assays
on every PCR Array
All PCR Arrays are Wet-Bench Validated
qPCR Assays on each PCR Array are tested for
Sensitivity:
constant C(t) value for same amount of template
Amplification Efficiency:
single curve analysis and standard curve dilutions
Specificity:
single: single melt-peak during dissociation
Validation performed on each lot (synthesized oligonucleotides) of Primers


- 22 -


PCR Arrays for All Biomedical Researchers
~ 100 different pathways for Human, Mouse, Rat, Monkey and Fly.
Cancer and Apoptosis

Cytokines & Inflammation

Development & Stem Cells

Apoptosis

Inflammatory Cytokines

Stem Cells

Cell Cycle

Th17 for Inflammation (NEW!)

Wnt Signaling

Human miRNA Array (NEW!)

Common Cytokines

Notch Signaling

Breast Cancer & Estrogen Receptor

Chemokines

TGFβ / BMP Signaling

Tumor Metastasis

NF-κB Signaling Pathway

Endothelial Cell Biology

Cancer PathwayFinder

Th1-Th2-Th3

Osteogenesis

Angiogenesis

TNF Ligands

Growth Factors

Cancer Drug Resistance

Toll-like Receptors

ECM & Adhesion

Signal Transduction

Toxicology & Drug Metabolism

Neuroscience

Signal Transduction PathwayFinder

Drug Metabolism

Neuroscience Ion Channels

NFkB Signaling

Drug Phase I Enzymes

Neurotransmitter Receptors

Jak / Stat Signaling

Drug Transporters

Neurotrophins & Receptors

DNA Damage Signaling

Oxidative Stress

Neurogenesis and Neural Stem Cell

Insulin Signaling

Stress & Toxicity

MAP Kinase Signaling

Other Diseases

Custom PCR Arrays

cAMP / Calcium Signaling

Atherosclerosis

96-Well Custom Arrays

p53 Signaling

Diabetes

384-Well Custom Arrays

Complete PCR Array List


- 23 -


How Genes on PCR Arrays Are Selected
Biologically relevant gene content

.

Not simply biochemical pathways or kinase cascades
Published association with the biological or disease
pathway
Specific gene information is gathered from a large
number of different overlapping sources including:
- Multiple Publicly Accessible Databases
- Text Mining Relevant Literature

Technically relevant gene content
Use genes that are regulated at the mRNA level
Specific feedback from thought leaders

.

Genes with High Relevance


- 24 -


Customizable PCR Arrays
Analyze the Genes Most Important
To Your Research

.

.

Available for any real-time PCR machine.
Wet-Bench Validated design and
performance.
2 to 3 week turn around time.
Same Data Analysis as Catalogued Arrays


- 25 -


PCR Arrays for All Biomedical Researchers
~ 100 different pathways for Human, Mouse, Rat, Monkey and Fly.

Cancer Research

Research Question:
What are the gene expression differences
between a metastatic and non-metastatic
Breast Tumor?

Angiogenesis
Epithelial to Mesenchymal Transition
Tumor Metastasis
Cancer Drug Resistance
p53 Signaling Pathway
Wnt Signaling Pathway
Cell Cycle
Breast Cancer and Estrogen Receptor Signaling
DNA Damage Signaling Pathway
MAP Kinase Signaling Pathway
Angiogenic Growth Factors and Inhibitors
Protein Phosphatases
PI3K-AKT Signaling Pathway


- 26 -


Application Example
Pathway Focused Analysis between metastatic and non-metastatic tumors

What is the gene expression differences between a
metastatic and non-metastatic Breast Tumor?
Tumor Metastasis PCR Array:
•
•
•
•
•
•
•
•
•
•
•
•
•

Cell Adhesion
Matrix Metalloproteinases
MMP Inhibitors
Cell Cycle Regulatory Genes
Cell Cycle Arrests and Checkpoint genes
Negative Regulators of Cell Proliferation
Positive Regulators of Cell Proliferation
Growth Factors and Hormones
Cytokines and Chemokines
Receptors
Pro-Apoptosis
Anti-Apoptosis

Run 3 PCR Arrays for each sample to calculate FC (and p value) using the ∆∆Ct method


- 27 -


Scatter Plot Result

∆
Log 10 2-∆C(t) Breast Tumor (metastatic)

UP REGULATED

MMP9
MMP2

FGFR4

MMP7

ETV4
MMP3
MMP10

DOWN REGULATED

∆
Log 10 2-∆C(t) Breast Tumor (non-metastatic)


- 28 -

Scatter Plot

MMP7
MMP9

MMP3
Log10 (P-value)

Statistical Significance

Volcano Plot Result

ETV4

MMP10

MMP2

FGFR4

DOWN REGULATED

UP REGULATED

Log 2 Fold Change


- 29 -

Volcano Plot

Seminar Overview

Cancer Research

Apoptosis


Cell Cycle

Inflammation

ECM/Adhesion

Neuroscience

Modifications

Signal Transduction

Stem Cell/
Development


Toxicology/
Drug ADME


- 30 -


What Is Micro RNA?

Endogenously expressed small functional RNAs (~ 21-23 nt)

.

Regulate mRNA expression post-transcriptionally
Primarily translation inhibition, but also mRNA degradation

.

Only partial sequence complementarity required
One miRNA can regulate multiple mRNA
One mRNA can be regulated by multiple miRNA

Another layer of complexity for regulating gene expression

.

.


- 31 -


miRNAs as Biomarkers in Cancer

Is there specific miRNAs expression differences in Human
Colon tumors versus normal adjacent tissue?
Normal Colon or Colon Tumor Tissue

Extraction of total RNA
QIAGEN
miRNeasy Mini Kit

miRNA


- 32 -

miScript miRNA
PCR ARRAYS


miRNA Biosynthetic Pathway

Mature miRNA
P
Dicer

miRNA

pre-miRNA

RISC

miRNA-RISC

Drosha

P

miRNA target recognition
pri-miRNA

Nucleus

Translation Inhibition
Message Turnover

miRNA gene


- 33 -


miRNA Biosynthetic Pathway

Mature miRNA
P
Dicer

miRNA

pre-miRNA

RISC

miRNA-RISC

Drosha

P

miRNA target recognition
pri-miRNA

Nucleus

Translation Inhibition
Message Turnover

miRNA gene


- 34 -


miScript miRNA PCR Arrays
cDNA Synthesis
1 hours

.

Load Plates (Preferably with 8-Channel
Pipettors)
2 minutes
.

2 hours

.

15 minutes

.


- 35 -


Anatomy of a catalogued miRNA PCR Array
miScript miRNA PCR Array Human miFinder (MIHS-001Z)

84 Pathway-Specific miRNAs

.

miR- miR- miR- miR- miR142-5p 16 142-3p 21
15a

miR29b

Let7a

miR126

miR143

Let7b

miR27a

Let7f

miR- miR9
26a

miR24

miR- miR30e 181a

miR- miR- miR29a 124 144

miR- miR30d 19b

miR- miR122
22

miR- miR150
32

miR- miR- miR155 140-5p 125b

miR- miR- miR141 92a 424

miR- miR17
191

miR- miR130a 20a

miR- miR27b 26b

miR146a

miR- miR200c 99a

miR- miR- miR19a 23a 30a

Let7i

Let7c

miR101

Let7g

miR425

miR- miR15b 28-5p

miR- miR- miR18a
25
23b

miR- miR302a 186

Let7d

miR- miR30c 181b

miR- miR223
320

miR374a

miRNA isolation control n=2

.

6 “Housekeeping”
snRNAs

.

miR93

miR106b

.

miR- miR- miR103
96
302b

miR- miR29c
7

Let- miR- miR- miR- miR- miR7e 151-5p 374b 196b 140-3p 100

miR- miR- miR- miR- miR194 125a-5p423-5p 376c 195

SNORD SNORD SNORD SNORD SNORD RNU
CelCel68
72
95
96A
miR-39 miR-39 61
6-2

miR- miR- miR- miR222 28-3p 128a 302c
MiRTC

MiRTC

PPC

PPC

miRNA Reverse Transcription
Controls (miRTC) n=2

.

Positive PCR Controls
(PPC) n=2

.

The miRNA PCR Arrays and reagents are intended for molecular biology applications. This product
is not intended for the diagnosis, prevention or treatment of disease.

- 36 -


miScript miRNA RNA PCR Arrays:
Available for Human, Mouse, Rat and Dog, Rhesus Macque
miRNome (miRBase V16)

Human

Mouse Rat Dog

miRNome (miRBase V16) 384 Well Format

Human

Mouse Rat Dog

miFinder

Human

Mouse Rat Dog Rhesus Macaque

miFinder 384HC

Human

Mouse

Brain Cancer

Human

Mouse Rat

Breast Cancer

Human

Mouse Rat

Cancer PathwayFinder

Human

Mouse Rat

Cell Differentiation & Development

Human

Mouse Rat

Immunopathology

Human

Mouse Rat

Inflammatory Response and Autoimmunity

Human

Mouse Rat

Neurological Development & Disease

Human

Mouse Rat

Ovarian Cancer

Human

Mouse Rat

Prostate Cancer

Human

Mouse Rat

Serum & Plasma

Human

Mouse Rat

T-Cell & B-Cell Activation

Human

Mouse Rat

miRNA QC

Human

Mouse Rat Dog


- 37 -


miRNAs as Biomarkers in Cancer


QIAGEN
miRNeasy Mini Kit

miRNA


- 38 -

miScript Human
CancerPathwayFinder
miRNA PCR Array


Colon Cancer Biomarkers

miScript Human CancerPathwayfinder PCR Array:
Many Cancer-Specific miRNA Are Up-regulated in the
Colon Tumor.


- 39 -


Colon Cancer Biomarkers

What about targets of the miRNAs?
Is it possible to measure mRNA from the same sample?


- 40 -


Use PCR Arrays to Measure Pathway Specific Gene Expression


QIAGEN
miRNeasy Mini Kit

RT2 Profiler
PCR ARRAYS
Archive “large” mRNA fraction
For use on PCR Arrays


miRNA

- 41 -

miScript
miRNA ARRAYS


Seminar Overview

Cancer Research

Apoptosis


Cell Cycle

Inflammation

ECM/Adhesion

Profiling Histone Modification on CDKN1A

Neuroscience

Signal Transduction

Stem Cell/
Development
Toxicology/
Drug ADME


- 42 -


CpG Islands and Methylation
•

CpG islands: CpG-rich regions in human gene promoters
–

Most CpG dinucleotides are methylated outside of promoter regions

–

Healthy/normal cells: rarely methylated

–

Cancer cells: hypermethylated, leading to transcriptional silencing

Normal

1

2

3

1

2

3

Expression

Promoter Region

Cancer
–

Important genes (i.e. tumor suppressors) are silenced

–

No Expression

All cellular pathways related to cancer can be affected

Adapted from Herman,
J. and Baylin, S. N. Engl.
J. Med. 2003


- 43 -


Background
DNA Methylation
Biological Importance
Role in normal development and also disease progression
Potentially silences critical tumor suppressors
Involves all cellular pathways
Stable/Covalent modificaton which can be detected in heterogeneous
fluids
Promising biomarker for early detection and predicting prognosis

.


- 44 -


DNA Methylation and Transcriptional Repression

Blood, 93:4059-4070, 1999


- 45 -


Methylation in Oncogenesis: Study Design
What are the difference in the Methylation status of breast cancer
associated genes among commonly used Breast Cancer Cell lines?

Obtained genomic DNA from Human Breast Cancer Cell lines (ATCC)
Normal Breast Tissue (Capital Bioscience)

Bioinformatics text mining:
Custom PCR Arrays:
genes hypermethylated in Breast Cancer with high frequencies


- 46 -


EpiTect Methy lI PCR Arrays (24 and 96 gene panels)
Simultaneously analyze the DNA methylation status of 22- or 94-gene panels associated with
specific cancer types, other diseases or biological pathways.
96 and 384 well qPCR Arrays-Human and Mouse
Cancer Related
Breast Cancer

Pathway-Focused Methyl qPCR Arrays
•
•
•
•
•
•
•
•
•
•
•
•
•

Epithelial to Mesenchymal
Transition
Gastric Cancer
Liver Cancer
Lung Cancer
Prostate Cancer
Colon Cancer
Leukemia & Lymphoma
Tumor Suppressor Genes

Apoptosis
DNA Repair
Stem Cell Transcription Factors
Inflammatory Response
T Cell Activation
Cytokine Production
Tumor Suppressor Genes
Homeobox (HOX) Genes
Polycomb (PcG) Genes
Cell Cycle
Toll-Like Receptor Signaling
TGF-Beta/BMP Signaling
Wnt Signaling

Custom Arrays and Custom Assays
Available for most CpG island-containing promoter in the human, mouse, or rat genome.
http://www.sabiosciences.com/dna_methylation_custom_PCRarray.php

- 47 -


DNA Methylation PCR Array Protocol Overview
(0.5 – 4 ug of Genomic DNA)

24 gene- Signature Panel


- 48 -





- 49 -



Data Analysis using downloadable Excel Templates
Accurate calculates ∆Ct values and converts to %



- 50 -


EpiTect Methyl II Biomarker Discovery

EpiTect Methtl II PCR Arrays can
be effectively used to identify
novel hypermethylated
biomarkers.


- 51 -


EpiTect Methyl II Biomarker Discovery
EpiTect Methtl II PCR Arrays can
be effectively used to identify
novel hypermethylated
biomarkers.

Measure gene
expression using
cataloged or custom
RT2 Profiler PCR Arrays
Use Epitect Pyrosequencing
Assays to quantitatively
identify Methylation at the
single nucleotide level.


- 52 -


Seminar Overview

Cancer Research

Apoptosis


Cell Cycle

Inflammation

ECM/Adhesion


Neuroscience

Signal Transduction


Stem Cell/
Development
Toxicology/
Drug ADME


- 53 -


What IS ChIP?

What is ChIP?
Chromatin Immuno-Precipitation
Method to examine in vivo protein-DNA interactions
Applications:
Transcription Factors, Modified Histones, Other nuclear factors
.

.

.

ChIP-qPCR is Chromatin Immunoprecipitation with qPCR Detection and Quantification


- 54 -


Applications for ChIP

Where does my Transcription Factor (or any protein that binds DNA) interact with
gDNA?
Example: Which p53 binding site is being used?
Use Custom ChIP PCR Array for multiple “tiles” within 1 gene
Or use individual ChIP qPCR Assay
What is the chromatin structure? Is it open or closed? (histone modifications)
Example: Can p53 bind to this site?
Use Cataloged Array for pathway (84 genes looking at +1 “tile”
Or Use individual qPCR Assay/ Custom PCR Array


- 55 -


ChIP PCR Arrays: 2 different applications and Layouts
Profiling 1 gene with 30 tiles
What is the chromatin structure for the entire promoter region

Profiling 1 tile (+1 tile) for 84 genes in a Pathway
•Tumor Suppressor & Oncogenes
•Cancer Drug Targets
•DNA Repair
•Cell Cycle
•Stem Cell Transcription Factors
•Inflammatory Response
•T Hellper Cell Differentiation
•Polycomb & Trithorax Target Genes
•Polycomb & Trithorax Complexes
•Homeobox (HOX) Genes


- 56 -


Chromatin Immunoprecipitation Workflow
Stimulus
X-Link
Lyse
Fragment

X

IP

X

X
Reverse X-Link
Purify DNA
Input

Anti-TF

NIS
Real-Time
PCR Primers

ChIP-qPCR


- 57 -


qPCR Assay Design: Tiling Scheme
TSS (+1)

Structural Gene
mRNA
gDNA

Distal Promoter
1 kb

Proximal Promoter
1 kb

.

1 kb

qPCR Assay & Amplicon

1 kb

1 kb

Amplicon Coverage Range (Tile)

Thirty (30) 1-kb tiles from -20 kb to +10 kb relative to each TSS
Accommodate every human, mouse and rat gene promoter
Balance Between Sensitivity & Resolution
Quality Controlled for Specificity & Amplification Efficiency

.

.

.

.


- 58 -


qPCR Assay Design: Tiling Scheme
for Pathway Catalogued Arrays
TSS (+1)

Structural Gene
mRNA
gDNA

Distal Promoter
1 kb

Proximal Promoter
1 kb

.

1 kb

qPCR Assay & Amplicon

1 kb

1 kb

Amplicon Coverage Range (Tile)

84 Genes with
only +1 Tile

Tiles +1kb relative to each TSS for 84 genes in a pathway
Quality Controlled for Specificity & Amplification Efficiency

.

.

Use the +1kb tile as a surrogate for promoter region to know transcriptional status
Use many genes to understand pathway
Correlate with gene expression

.

.

.


- 59 -


Modified Histone Distribution Across a Promoter

Custom ChIP PCR Array for Entire CDKN1A Gene
Histone markers for actively transcribed genes (H3Ac and H3K4me2) but not for
transcriptionally inactive genes (H3K27me3) surround the CDKN1A gene making it
available to p53 regulation.

- 60 -


Seminar Overview

Cancer Research

Apoptosis


Cell Cycle

Inflammation

ECM/Adhesion


Neuroscience

Signal Transduction


Stem Cell/
Development
Toxicology/
Drug ADME


- 61 -


PCR Array Summary
Complete System (From Sample to Fold Change)
Five-component system: Sample Isolation, PCR arrays, First strand
(OR DNA RE Kit), master mix, and data analysis software

.

Breadth of Content CATALOGED Pathways or customizable
Gene Expression
MicroRNA Expression
DNA Methylation
Chromatin Immunoprecipitation (histone modifications/TF binding)
Validated Real-Time PCR Performance
Wet-Bench tested qPCR Assays for Sensitivity, Specificity, and
Amplification Efficiency on every qPCR manufactured.
Final Benefit to Researcher:

.

.

Platform technology for any qPCR machine in any laboratory

- 62 -


Need More Info? Product Support Tools

www.sabiosciences.com
Global Technical Support Email: Support@SABiosciences.com

- 63 -


Thank you for attending!

Questions, Comments, Suggestions, Concerns?

Samuel.Rulli@QIAGEN.com
US Applications Support
support@sabiosciences.com


Global Applications Support
SABio@qiagen.com

- 64 -


Understanding Gene Expression and Epigenetics Using PCR Arrays

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