abclt EASY TO USE MICROARRAY PRODUCTS-TODAY! Pat Mayer Global Product Manager MICROMAX Feb. 2002.

EASY TO USE MICROARRAY PRODUCTS-TODAY!EASY TO USE MICROARRAY PRODUCTS-TODAY!

Pat MayerGlobal Product ManagerMICROMAX Feb. 2002

PerkinElmer Microarray SolutionPerkinElmer Microarray Solution

Scanners

Complete Microarray Systems

GeneTAC Hybstation

Labeling Kits

NEN Brand Nucleotide Analogs (Cyanine- and Biotin-labeled)

Prespotted Microarrays

Glass Slides

Spotters

Data Analysis Software

PerkinElmPerkinElmerer

GenomicsGenomics

GeneTAC G3

Slide Scanning Service

PerkinElmer Microarray SolutionPerkinElmer Microarray Solution

Why MICROMAX Products?Why MICROMAX Products?

Why MICROMAX Products?Why MICROMAX Products?

Differential gene expressionDifferential gene expression

Gene expression

Relative levels of RNA and proteins

Differential gene expression

Differences in levels of RNA and proteins as a result of changes

in cellular phenotype

Backgound MICROMAX TechnologyBackgound MICROMAX Technology


MicroarraysMicroarrays

A new and prominent technology that vies with A new and prominent technology that vies with PCR as the most significant commercial scientific PCR as the most significant commercial scientific advance of the decade.advance of the decade.

Technology allows

Miniaturized formatMembrane or glass slideDetection instrumentationAnalysis software


DNA MicroarraysDNA Microarrays

Membrane Arrays

Oligonucleotide Arrays

cDNA or Long Oligo Arrays


Types of DNA ArraysTypes of DNA ArraysMembrane Arrays

cDNA spotted on nylon membraneRad nucleotides (chemiluminesence)Gene expression and novel gene discovery

Advantages Standard molecular biology techniquesSensitiveReusableUnknown genes (ESTs) can be analyzed

Disadvantages Single analysis per arrayLow densityRadioactive safety


Oligonucleotide chips

Oligo probes synthesized onto chipFluorescence detectionGene expression and mutational analysis

Types of DNA ArraysTypes of DNA Arrays

Advantages Very specific, can identify isotypic changes or SNPsTens of thousands oligos can be analyzed per chipAutomation

DisadvantagesOnly known genes can be analyzedSingle analysis per arrayLow sensitivityMany spots make a geneMay be too specific for gene expression analysisNot reusable


Glass microarrays

cDNA or long oligos (50-70 mers) printed onto slidesTwo-color fluorescence detectionGene expression and novel gene discovery

Advantages Dual detection for accurate differential analysisThousands of individual cDNA per slideUnknown genes (ESTs) can be analyzedAutomation

Disadvantages Low sensitivityNot reusable

Types of DNA ArraysTypes of DNA Arrays


Labeling and detection for MicroarraysLabeling and detection for Microarrays

Direct - radioactivity

Reverse Transcriptase

33P dNTPmRNA

Preferred method for membrane arrays

** Single detection , Very sensitive**

cDNA

*P* *P**P*


Labeling and Detection for MicroarraysLabeling and Detection for Microarrays

Direct - fluorescence


fluoro- dNTPmRNA

Standard method for glass and oligo arrays

** Dual detection, Good sensitivity**

cDNA

F F F


Labeling and Detection forLabeling and Detection for MICROMAXMICROMAX

Indirect and amplified - Fluorescence


biotin-dNTPfluorescein-dNTP

mRNA

Excellent method for glass arrays (MICROMAX-TSA)

** Dual detection, Excellent sensitivity**

YH YH YHTF T

F

TF

TF

TF

T F

TF

TF

TF

TF

cDNA

B B B

TSA Technology ReviewTSA Technology Review


TSA is a proprietary, PATENTED technology developed at NEN

Patented under the name CARD (Catalyzed Reporter Deposition)

Commercialized under the name TSA (Tyramide Signal Amplification)


Inroduction of Horseradish Peroxidase (HRP) enzyme

Keys To TSA

Labeled Tyramide


SIGNAL AMPLIFICATION




SENSITIVITY

SENSITIVITY

SENSITIVITY

Signal DetectionSignal Detection

Used existing patented and proprietary technologyUsed existing patented and proprietary technology

TSATyramide Signal Amplification

Sample immobilized to a surface - slide

B B BSpecific binding with

hapten containing probe

YH YH YHIncubate with HRP- conjugate

TF

TF

TF

React with Tyramides

TF

TF

T F

TF

TF

TF

TF

TF

TFTF

Amplify

TSA on MicroarraysTSA on Microarrays

Sequential Fluorescence Deposition with TSASimultaneous hybridization

both Fluorescein and Biotinlabeled total RNA

Incubate with streptavidin-HRP

Amplify with TSA-cyanine 5

Incubate with Anti-F-HRP

YYYY

YYYY

Amplify with TSA-cyanine 3

Inactivate Residual HRP


Reduced Customer Sample RequirementsTSA reduces the input RNA sample up to 200-fold

Increases opportunity for microarray technology useTumor samplesReduced RNA sample costsIncreased # of experiments

MICROMAX microarray Analysis with Direct Fluorescence MICROMAX microarray Analysis with Direct Fluorescence and Tyramide Signal Amplificationand Tyramide Signal Amplification

Direct Labeling Method TSA Labeling Method

4800 Slide

(100 µg Total RNA) (1 µ g Total RNA)

TSA labeling method is very sensitive and needs as little as 0.5 to 1 µ g total RNA

Direct and TSA Labeling: Scatter PlotsDirect and TSA Labeling: Scatter Plots

1 10 100 1000 10000

SA177_Direct_Cy5_P67_L80 data.gpr (control)0.01

0.1

1

10

100

1000

10000

1 10 100 1000 10000

SA197_TSA_Cy5_P65_L45 data.gpr (control)0.01

0.1

1

10

100

1000

10000

HL-60: Cyanine 5

Jurk

at:

Cya

nin

e 3

Jurk

at:

Cya

nin

e 3

HL-60: Cyanine 5

Direct TSA

TSA labeling method is highly sensitive and identifies more differentially regulated genes

Fully integrated systems for streamlined application and consistent performance.

MICROMAX Platforms

tissue samples isolatetotal RNA

cDNAlabeling

Biotin/Fluorescein linkeddNTPs

Cyanine 3/5dNTPs OR

TSATM amplified fluorescence detection

hybridize hybridize

imagingdata analysis

MICROMAX Products and Services MICROMAX Products and Services == FlexibilityFlexibility

Slide Scanning and DataProcessing Services

CompleteMicroarray Systems

MICROMAXTM DirectMicroarray Systems

MICROMAXTM TSAMicroarray Systems

Human Array Systems I&II

Chemical Labeling Kit

Microarray Products

Cyanine 3- labeled dCTP, dUTP

Cyanine 5- labeled dCTP, dUTP

Human Screening microarrays

Human Focused microarrays

Quality Control microarray (Practice Slide: 25 genes)

Reagent Kits

Direct KitTSA Kit

Human Focused Array Systems

Glass SlidesULTRACLEAN ISUPERCHIP I

Human Screening Array System I&II

MICROMAX DirectMICROARRAY SLIDES

2 human cDNAmicroarrays

2 practice arrayscoverslips

cDNA PREPARATIONCyanine 3-dUTPCyanine 5-dUTPReverse Transcriptase

(RT)/RNase inhibitordNTP labeling mixUnlabeled control RNA

HYBRIDIZATIONHybridization Buffer

INSTRUCTION MANUAL

System ComponentsSystem Components

MICROMAX TSA

MICROARRAY SLIDES2 human cDNA

microarrays2 practice arrayscoverslips

cDNA PREPARATIONBiotin and Fluorescein Nucleotides

dNTP Nucleotide MixReverse Transcriptase

(RT)RNase InhibitorUnlabeled control RNA

cDNA QC CHECK AND HYBRIDIZATIONBiotin- and Fluorescein-Control cDNAsGeneScreenTM MembraneStreptavidin-HRP and Anti-Fluor HRPHybridization Buffer

DUAL FLUORESCENCE DETECTIONTSA Amplification BufferCyanine 3-TyramideCyanine 5-TyramideHRP Inactivation Reagent

System ComponentsSystem Components

Platform SpecificationsPlatform Specifications

Sample RequirementSample Requirement• 20-80 ug Total RNA20-80 ug Total RNA

Dynamic RangeDynamic Range• 8-125,000 pg input mRNA 8-125,000 pg input mRNA

(>10(>1044)) Differential SensitivityDifferential Sensitivity

2-fold ratio difference2-fold ratio difference

Sample RequirementSample Requirement• 0.5-1 ug Total RNA0.5-1 ug Total RNA

Dynamic RangeDynamic Range• 0.04-200 pg input mRNA 0.04-200 pg input mRNA

(>10(>103.53.5)) Differential SensitivityDifferential Sensitivity

3-fold ratio difference3-fold ratio difference

MICROMAX DirectMICROMAX Direct MICROMAX TSAMICROMAX TSA

MICROMAX Products & ServicesMICROMAX Products & Services

MICROMAX Human cDNA Microarray System I-DirectDirect Fluorescence Labeling System

5 slides

MICROMAX Human cDNA Microarray System 1.1-TSATSA Fluorescence Labeling System

5 slides

MICROMAX Human cDNA Microarray System II-TSATSA Fluorescence Labeling System

5 slides

PositioningEverything for novice microarray user

Complete MICROMAX Systems


MICROMAX Direct Reagent KitDirect Fluorescence Labeling Kit

10 Labeling Reactions50 Labeling Reactions

MICROMAX TSA Labeling and Detection KitTSA Fluorescence Labeling Kit

10 Labeling & Amplifcation Reactions40 Labeling & Amplifcation Reactions

MICROMAX Reagent Kits

PositioningResearchers who manufacture their own microarrays


MICROMAX Pre-Spotted Microarrays

MICROMAX Microarray: Human cDNA 1.1 (2400 human genes)MICROMAX Microarray: Human cDNA II (4800 human genes)MICROMAX Microarray: Quality Control SlidesMICROMAX Microarray: Focused Array Slides (~270 human genes)

2 Microarrays

PositioningExperienced researchers who don’t have access to cDNA arrays

Experienced researchers who have their own labelingtechnology

GeneMAP Microarrays: Human, Mouse and Rat

5 microarrays


MICROMAX Slide Scanning Service

Scanning of all types of MICROMAX SlidesPractice Arrays free of charge

Slide Scanning Service UpgradedWeb Based Service

PositioningNovice researchers who don’t have access to fluorescent microarray scanner

Selling GeneMAP MicroarraysSelling GeneMAP Microarrays

What are they?What are they?• Spotted, preprinted arraysSpotted, preprinted arrays• Human, mouse and rat glass microarraysHuman, mouse and rat glass microarrays

Available Worldwide (non-exclusive) from Available Worldwide (non-exclusive) from PerkinElmerPerkinElmer

Sales of GeneMAP will count towards your MM sales Sales of GeneMAP will count towards your MM sales goals!goals!• Under a different pac code however, 2063Under a different pac code however, 2063

MICROMAX CompetitionMICROMAX Competition

MM


Who sells “Complete Systems”?Labeling SystemNucleotidesSlides

ONLY PERKINELMER!


CompetitionCompetition

Company Labeling Kits Slides

Affymetrix x X

Agilent XClontech X XAmersham XStratagene X XGenisphere XMergen X

HOMEBREW!!!! X X


cDNA Labeling Technology

AminoAllylClontech

Dendrimer Genisphere

Slide Technology

Short (20-30 mers) OligoAffymetrixMergen

Medium (70-80 mers) OligosClontech

cDNAAgilentTakara

Competing Technologies

HOMEBREW

HOMEBREW

MICROMAX Competition-TechnologiesMICROMAX Competition-Technologies

Amino groups are incorporated during cDNA synthesis using aminoallyl-dUTP

Cy3 or Cy5 monomer NHS ester is reacted with cDNA

Labeling is independent of incorporation of dye-dNTP analogs

Aminoallyl labeling technology…….

……is a novel technology which is cumbersome

TARGET CUSTOMER FOR NEW CHEMICAL LABELING!

MICROMAX Competition-TechnologiesMICROMAX Competition-Technologies

Labeling of cDNA RT with Unlabeled deoxynucleotides and Cy3™ or Cy5™ Primer Oligo (=3DNA Capture Sequence)

Hybridize to Array

Detect with 3DNA Fluor (Complementary Capture Sequence labeled with Cy-Dye™)

…...only competing technology to MICROMAX-TSAOur customers have told us this doesn’t work very well-no specific signal

Genisphere’ s dendrimer technology…...

Competitive AdvantagesCompetitive Advantages

Low cost entry to microarray technologyLow cost entry to microarray technology

Complete systems right out of the boxComplete systems right out of the box

Broad product offeringBroad product offering

License to do dual color fluorescence hybridizationLicense to do dual color fluorescence hybridization

Every lab can afford it!Every lab can afford it!

Easy to use!Easy to use!

Something for every lab!Something for every lab!

Two samples on one slide!Two samples on one slide!

MICROMAX SellingMICROMAX Selling

Are you currently using microarray technology in your research?Are you currently using microarray technology in your research?

Do you anticipate using microarray technology in the near future?Do you anticipate using microarray technology in the near future?

Is your differential gene expression research limited by the amountIs your differential gene expression research limited by the amountof starting material?of starting material?

Are you just getting started in microarray technology?Are you just getting started in microarray technology?

Probing Questions…...

…….leads to providing customers with the right products

Labeling and Detection KitsLabeling and Detection Kits

• QC all componentsQC all components

• Nucleotides HPLC QCNucleotides HPLC QC

• Nucleotides functional Nucleotides functional testingtesting

• Components used in Components used in functional testing of functional testing of microarraysmicroarrays

• Cross check in R & D and Cross check in R & D and Applications LabApplications Lab

MicroarraysMicroarrays

• Check PCR processCheck PCR process

– In process checksIn process checks

• Check autofluorescence of Check autofluorescence of glass slidesglass slides

• Representative sequencingRepresentative sequencing

• Check after printing for spotsCheck after printing for spots

• Functional testing of arraysFunctional testing of arrays

– Spot morphologySpot morphology

– Spot sizeSpot size

– S/NS/N

• Cross check in R & D and Cross check in R & D and Applications LabApplications Lab

QC Process for Microarray productsQC Process for Microarray products

New MICROMAX products in the last yearNew MICROMAX products in the last year

4800 Arrays and Systems (1.1) (Alphagene genes)4800 Arrays and Systems (1.1) (Alphagene genes)• Now manufacture our own arraysNow manufacture our own arrays• TSA and Direct versionsTSA and Direct versions

Direct Kit Direct Kit • more economical-$40/slidemore economical-$40/slide

Focused ArraysFocused Arrays• Human Apoptosis & NeurobiologyHuman Apoptosis & Neurobiology

Additional Screening Arrays and System (II)Additional Screening Arrays and System (II)• Incyte Genomics’ Gene ContentIncyte Genomics’ Gene Content

Coming SoonComing Soon• Glass Slides for DNA spottingGlass Slides for DNA spotting• New Chemical Direct Labeling SystemNew Chemical Direct Labeling System• Mouse and Rat Screening ArraysMouse and Rat Screening Arrays

– Content from LION bioscienceContent from LION bioscience• Control KitsControl Kits

NEW

Current MICROMAX AdsCurrent MICROMAX Ads MICROMAX TSAMICROMAX TSA

• ““The NEN TSA Microarray Labeling and Detection Kit lets you do more with less”The NEN TSA Microarray Labeling and Detection Kit lets you do more with less”• Ran about 10 times in 2001-will be redesignedRan about 10 times in 2001-will be redesigned

MICROMAX Direct Kit AdMICROMAX Direct Kit Ad• ““Simply the best microarray reagents on earth (or anywhere else you would care to Simply the best microarray reagents on earth (or anywhere else you would care to

explore!”explore!”• Ran about 2 times in 2001 and continue in 2002Ran about 2 times in 2001 and continue in 2002

MICROMAX Microarray AdMICROMAX Microarray Ad• ““A long time ago. . .in a galaxy far away. . .we started making pre-spotted A long time ago. . .in a galaxy far away. . .we started making pre-spotted

microarrays. . .We Still Do.”microarrays. . .We Still Do.”• Newly created and will run in 2002Newly created and will run in 2002

MICROMAX Glass Slide AdMICROMAX Glass Slide Ad• ““We have a new addition to the family. . .Introducing MICROMAX Glass Slides”We have a new addition to the family. . .Introducing MICROMAX Glass Slides”• Newly created and will run in 2002Newly created and will run in 2002

MICROMAX Chemical Direct Labeling AdMICROMAX Chemical Direct Labeling Ad• Not created yet and will run in 2002Not created yet and will run in 2002

Introducing MICROMAX Glass SlidesIntroducing MICROMAX Glass Slides

ULTRACLEAN IULTRACLEAN I• Glass Slides that are cleaned!Glass Slides that are cleaned!• For homebrew coating proceduresFor homebrew coating procedures

– Looking for evaluators (e-mail Pat)Looking for evaluators (e-mail Pat)– MPS692MPS692

SUPERCHIP ISUPERCHIP I• Amino Coated Glass SlidesAmino Coated Glass Slides

– Looking for evaluators (e-mail Pat)Looking for evaluators (e-mail Pat)– MPS695-5 slidesMPS695-5 slides– MPS696-20 slidesMPS696-20 slides

Bundling opportunities with PerkinElmer printers and spotters!Packaged in sets of 5 slidesReady to ship evals February 11, 2002Ready to ship by end of February

Ultra-flatUltra-flatLow Intrinsic FluorescenceLow Intrinsic Fluorescence

Breadth of Technical SupportBreadth of Technical Support

In-field dedicated genomics specialistsIn-field dedicated genomics specialists

Internal dedicated Technical SupportInternal dedicated Technical Support• Leslie Stern X9117Leslie Stern X9117

Internal dedicated Applications LabInternal dedicated Applications Lab• Testing and expanding microarray protocols Testing and expanding microarray protocols

Research and Product Development StaffResearch and Product Development Staff• Organized for efficiency and time to market improvementsOrganized for efficiency and time to market improvements• External seminar seriesExternal seminar series

MICROMAX Sales Tools AvailableMICROMAX Sales Tools Available

MICROMAX SeminarMICROMAX Seminar

Will be updated ASAP to incorporate Packard instrumentsWill be updated ASAP to incorporate Packard instruments

MICROMAX Literature PiecesMICROMAX Literature Pieces

CodeCode Name of PieceName of Piece

H78255H78255 MICROMAX Sell SheetMICROMAX Sell SheetH78511H78511 MICROMAX TSA BioTechniques ReprintMICROMAX TSA BioTechniques ReprintP10208P10208 MICROMAX Direct Reagent Kit Sell SheetMICROMAX Direct Reagent Kit Sell SheetH78414H78414 MICROMAX TSA Sell SheetMICROMAX TSA Sell SheetH78381H78381 MICROMAX Focused Array Sell SheetMICROMAX Focused Array Sell SheetH78416H78416 MICROMAX TSA Protocol Laminated CardMICROMAX TSA Protocol Laminated Card

Laminated card (H78416) is a must have for wet labs!Laminated card (H78416) is a must have for wet labs!

Order at www.perkinelmerliterature.com

abclt EASY TO USE MICROARRAY PRODUCTS-TODAY! Pat Mayer Global Product Manager MICROMAX Feb. 2002.

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Transcript of abclt EASY TO USE MICROARRAY PRODUCTS-TODAY! Pat Mayer Global Product Manager MICROMAX Feb. 2002.