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384 well PCR Cleanup Application

Launch package

January 24, 2003

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Page 2

Index

Technology Page

Overview PCR 3

Overview PCR Cleanup 4

Whatman Protocol 5

Protocol Automation 6

Product

System Components 7

Pricing/Packaging 8

Competitor Comparison

Whatman Performance Data 9

Performance Comparison 10Protocol Time Comparison 11

Value Page

Features/Benefits 12

Customer

Customer ID 13

Market Sizing 14

Product Positioning 15

Marketing Support

Sample Information 16

Advertising Plan 17

Trade Show Plan 18

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Page 3

PCR Overview

Capable of greater than a Billion

fold replication of a DNA sequence

of interest

Necessary to create enough DNA to

perform:

Hybridization assays

(i.e. microarray)

Sequencing

SNP detection methods

Genotyping

cDNA library construction

PCR has been the most powerful

and widely used molecular biology

technique for the past 30 years

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Page 5

Whatman 384 PCR Cleanup Procedure

Add Binding Buffer

PCR Product (5-30ul)

384 Well DNABinding Plate

2 x Wash with

90ul wash buffer

X 384

Add Elution Buffer

(10ul-50ul)

Centrifuge Filter

2.5 min @ 1800xg

Bind

Centrifuge Filter

5 min @ 1800xg

Wash

Elute

Centrifuge Collect

2.5 min @ 1800xg

Incubate

(1 min @ RT)

384 WellCollection Plate Collect

Waste - 384 Deep WellCollection Plate

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Page 6

Protocol Automation

Demonstrated adaptability shown with

an in-house automated liquid handler(Biomek 2000)

Adaptation to other laboratory

instrumentation will be case specific

(no universal solution)

SBS compatible platesThings to consider:

Pipetting speeds - Semi-viscous buffers require careful pipetting during

delivery, mix & transfer steps

Disposables - Customer specific PCR tubes & pipette tips will have an

effect on liquid handling parameters (pipetting heights/speeds, mixing& transfer steps)

Molecular Biology Precautions - Routine cleaning of processing

equipment and lab surfaces will reduce the risk of unwanted amplicon

contamination. Use of aeresol resistant pipette tips is recommended.

Biomek 2000

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System Components

Upstream Accessories

96-well PCR Plates (7705-1901)

384-well PCR Plates (7705-1305)

Heat seals (7704-0002 or 7704-0003)

Downstream 384-Well Accessories

384-well black polystyrene plates (7701-2100) - fluorescent measurements

384-well white polystyrene plates (7701-3100) - chemiluminescent measurements

384-well 0.45u hydrophilic PVDF filterplate (7700-2106) - media support plate forcustomer specific dye terminator cleanup

384-well PCR Cleanup Components

384-well DNA Binding Plate (7700-2110)

384-well 400ul Waste Collection Plate (7701-5400)

384-well 100ul Collection Plate (7701-1100)Adhesive plate seals (7704-0001)

Equipment required

Centrifuge with microplate holder (2.3 inch stack height)

Automated Liquid Handling System & associated disposables

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Page 8

Pricing

Package Weights

7700-2110 384-well DNA Binding Plate 50 790

7701-5400 384-well 400ul Waste Collection Plate 25 143

7701-1100 384-well 100ul Collection Plate 50 110

7704-0001 Adhesive plate seals 100 44

List Price/Prep 0.06

Cat # Description Height cm Weight k Length cm Width cm

7700-2110 DNA Binding Plate 13 4.31 43.18 14.24

7701-1100 Uniplt 384 Well 100ul 14 2.86 44.45 17.78

7701-5400 Uniplt 384 Well 400 ul 14 2.86 44.45 17.78

7704-0001 Adhesive Plate Seal 5.08 0.34 20.32 11.43

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Page 9

Pricing in Euros

Package Weights

Cat Num Description Plates/Case Price/Case

7700-2110 384-well DNA Binding Plate 50 1,304

7701-5400 384-well 400ul Waste Collection Plate 25 240

7701-1100 384-well 100ul Collection Plate 50 182

7704-0001 Adhesive plate seals 100 73

List Price/Prep E 0.06

Cat # Description Height cm Weight k Length cm Width cm

7700-2110 DNA Binding Plate 13 4.31 43.18 14.24

7701-1100 Uniplt 384 Well 100ul 14 2.86 44.45 17.78

7701-5400 Uniplt 384 Well 400 ul 14 2.86 44.45 17.78

7704-0001 Adhesive Plate Seal 5.08 0.34 20.32 11.43

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Page 10

Performance DataCapillary Sequencing

Purified 1.8kb product sequenced

using big dye chemistry

Non-cross talk

Alternating positive & negative cleanups re-amplified to

detect low levels of cross contamination

Development Run - Yield / Recovery / Reproducibility

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

4.00

4.50

5.00

213bp A melogeni n 500bp pGE M 1. 9k b pG EM

PCR Product

DNAQuantity(ug/w

ell)

86.0%

88.0%

90.0%

92.0%

94.0%

96.0%

98.0%

100.0%

%Recovery

Crude Input (ug)

DNA Yield (ug)

% Recovery

7.36%

CV

5.15%

CV

7.41%

CV

DNA Recovery & Consistency

3 Different PCR product sizes purified &

quantified (n=24)

384 well PCR Cleanup - Adjunct Study

0

1

2

3

4

5

6

7

8

9

10

0 2 4 6 8 10

HS DNA Input (ug/wel l)

DNAYield(ug/we

ll)

Standard PCR

2% DMSO

1.5M Betaine

0.1% Glycerol

DNA Binding Capacity

Varying levels of DNA Input purified and

quantified. Study also shows comparison of

PCR reactions containing typical adjucts

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Competitor Performance Comparison

Parameter Whatman Millipore Telechem=CPG=Genetix

Sample requirements 5-30ul PCR reaction (split

or concentrate if needed)

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Liquid handling pipette headMethod #

plates

8 channel* 384 channel**

2 1:25:10 21:30Whatman

4 2:36:50 29:30

Millipore 1 51:00 28:002 1:33:30 25:30CPG/Telechem/

Genetix 4 2:51:30 35:30*based on internal Biomek 2000 testing

**estimated times

Competitor Time Comparison

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Page 13

Features384 well

Bind-wash-elute chemistry

Centrifuge processing protocol

SBS compatible microplates

Sample volumes 5-30ul

High recovery of low MW PCR

products

Robust to various PCR adjuncts

Uses DNA Binding plate

Bulk Component Purchase

Benefits Increased throughput, decreased cost

Simple method

No phenol / chloroform / alcohol extractions

Provides highly purified DNA.

Easy downstream sample retrieval.

Complete elimination of well to well

contamination. Efficient volume recovery

Compatible with automated liquid handling

equipment

No sample concentration or dilution needed for

typical 384 PCR samples

Able to meet the demands of genomics customers

Compatible with lab specific PCR chemistries

Adaptable to binding nucleic acid from many

sample sources

Economic benefit for large genome centers and

custom kit manufacturers

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Page 14

Target Customers

Genomics OEM PotentialsKit companies: Qiagen

Amersham BioRad

Promega

Gentra

Genetix

Eppendorf

Applied Biosystems

Stratagene

Mackerey Nagle

Invitrogen Roche

Edge

Princeton Separations

Instrument companies: Perkin Elmer

Tecan

Beckman

Gilson

Zymark Brandel

Genomic InstitutionsUS: Albert Einstein College of Medicine

Baylor College of Medicine The Chromosome 21 Mapping and

Sequencing Consortium

Center for Genetics in Medicine

Genoscope

GTC Sequencing Center

Cold Spring Harbor

Joint Genome Institute, U.S.

Department of Energy

Stanford DNA Sequencing AndTechnology Development Center

Stanford Human Genome Center

The Institute for Genome Research

University of Utah Genome Center

University of Oklahoma Advanced

Genome Technology Center

University of Texas, Southwestern

Medical Center

University of Washington GenomeCenter

The Institute for Systems Biology

Whitehead Institute for Biomedical

Research

Washington University

Europe: Gesellschaft fur

Biotechnologische ForschungmbH

InstituteforMolecular

Biotechnology, Jena, Germany

Max Planck Institute for

Molecular Genetics

Sanger Center

Finland Genome Research

Center

Asia: Human Genome Center, Institute

of Genetics, Chinese Academy of

Sciences

National YangMing University

Genome Research Ceter

Japan: Keio Uiversity School of

Medicine, Tokyo

Japan Science and Technology

Corporation

RIKEN Genome Sciences Center

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Page 15

Target Market / Sizing

WW Market Size for PCR Clean up Kits ($M)

0.0

5.0

10.0

15.0

20.0

25.0

30.0

2000 2001 2002 2003 2004 2005 2006 2007

Sample Capacity of Thermal Cyclers Acquired 1998-2000

(Source PhorTech Thermocycler Mkt Survey)

0

10

20

3040

50

60

70

384 96 48 24

Sample Capacity

%o

fTHermalCyclersin

Installed

Base

The WW market for PCRClean up kits is estimated

at $15M in 2003 with

forecasted growth rate of

9% per year.

The shift to 384 well

PCR plates, starting in

2000, is gainingmomentum and leading

to demand for 384 well

PCR clean up.

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Page 16

Product Positioning

The 384 well PCR Clean up product is intended for use in

large genomic research institutions performing millions ofPCR reactions per year.

Bulk Product

Primary market research suggests that these large genome

centers will fine tune their own protocol and set up their own

automation systems, thereby requiring bulk components as

opposed to turnkey kits. Specific examples of this preference

include Sanger Center, Cold Spring Harbor and the Finland

Genome Research Center.

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Samples & Inventory

Sample plates and accessories can be ordered separately

Or as a 2 plate set

Cat Num Description Units/Case

s7700-2110 384-well DNA Binding Plate 1 plate

s7701-5400 384-well 400ul Waste Collection Plate 1 plate

s7701-1100 384-well 100ul Collection Plate 1 plate

s7704-0001 Adhesive plate seals 1 seals

Cat Num Description Units/Case

s7905-0004 384-well PCR Clean up sample pack 768 reactions

The protocol and buffer formulations are available at the

Whatman web site locations

Technical Support/Protocols/ApplicationsProtocols

Technical Support/Applications/Laboratory Filtration-Microplates/Technical Papers