`
`Some Like It Hot: A Thermal Cycler Roundup
`
`Former high-endfearures are standard
`options in the newest thermal Cyclers
`Bv AILIEH Consrmrs
`
`lids featuring automalic
`pressure control
`for
`reli~
`able sealing and protection
`of tubes.
`
`
`
`Brinkmann's Eppendorf Maflercycler line of thermal
`cycle-rs includes gradient and personal-sized model's.
`
`
`
`CourtesyuiBrinkmannInstruments
`
`It was bound to happen. During the growth of PCR,
`
`many companies joined the thermal cycler bandwagon.
`But since The Scientist's last [henna] cycler review.I
`some of these manufacturers—such as Ericomp—have left
`the cut-throat market and a number of the machines listed
`in previous LabConsumer profiles have been discontinued.
`Market niches remain for those companies and product
`lines that have Survived the intense competition.
`One such niche is real-time lherrnoeyclers. previously
`reviewed in The Scientist} These cyclcrs permit
`researchers to monitor the thermal and optical properties
`of their reactions in real time. and are indispensable for
`quantitative PCR. Another specialty is high-throughput
`PCR. spurred by the rapid growth of genomicsrrelated
`research. Overall. features that were once optional. such
`as heated lids. are new standard. and interchangeable
`blocks allow researchers to customize their experiments
`even with low«end thermal cycler models.
`Almuugh the number of companies manufacturing
`thermal cyclcrs may be on the decline. the number of
`options available to researchers is not. From basic labo—
`ratory PCR workhorses to real-time and highethroughput
`machines. the latest batch of thermal cyclcrs will certain-
`ly heat
`things up in the lab this coming year. What
`follows is an alphabetical
`round up of themmcycler
`providers:
`
`Applied Biosystems: Foster City. Calif-bascd Applied
`Biosystems holds the patents on thermal cyclets. along
`with the exclusive license to the PCR process,3 and offers
`researchers several PCR options. The GEDBAITIP® PCR
`System 9100 is designed for high sample volumes and
`comes in 60~welli05 ml. 384-well. and 96-well formats.
`The GeneAmp PCR System 2400 is a personal—sized
`cycle-r that features a heated lid and a 24-well sample
`block. The GeneAmp PCR System 2700 includes many
`of the features of the 9700 system but
`in a smaller
`footprint. For
`the real-time PCR market. Applied
`Biosystcms' AB] PRISM® 7700 Sequence Detection
`System features a built-in thermal cycler and the ability
`to detect multiple fluorophores in a single tube.
`
`Biometra: Goningen. Germany-based Biometm offers a
`variety of Poitier-based cyclcrs. ranging from personal-
`sizcd to high throughput. (The Pelticr effect transfers
`heat from one side of a semiconductor to another, creat-
`ing an efficient heating and cooling system without
`refrigerants or compressors] The Tpersorrnl rhen'nocy-
`cler
`features three different block formats—48mph.
`20-well. and combi-block. Each format handles tubes.
`strips. and microplates. The TGmdiem thermocycler
`allows fast optimization of new reactions with a thermal
`gradient of up to 40°C. The Tl thermocycler offers a
`heated l‘rd. gold-plated silver block. and heating and cool
`ing rates of 4°C per second and 3°C per second.
`respectively. The T3 thermocycler, the successor to the
`company's TRIO model. offers a choice of thrce formats:
`3 x 48 wells. 3 x 20 wells. and a combi-blocit with 3 x 18
`and 3 it 43 wells. The T3 also features three independent
`blocks with adjustable heated lids. allowing the user to
`run experiments with different sample sizes and formats
`in the same instrument. The TROBOTthermocyclcr is the
`newest addition to Biometra's cyclcr range. It is compat-
`ible with a variety of robotic systems and offers a
`motorized heated lid and a small lbutprint. The motor-
`ized plate lifter ensures easy removal of plates by a
`robotic am. All Biometra thermal cyclcrs have heated
`
`
`32 The Scientist
`December 10. 200}
`
`Bio-Rad Laboratories:
`Hercules.
`Calif.~based
`Bio~Rad
`offers
`the
`iCyclerTM Thermal Cyclc.
`a flexible. modular system
`that easily upgrades to realv
`time
`quantitative
`PCR
`capability and also offers
`optional gradienl capability to
`facilitate reagent optimization.
`The optical
`real—time detection
`system upgrade is a modular unit
`that installs directly over an existing or new ECycler unit.
`The rewiring iCycler iQ system is designed as an open
`platform. capable of detecting a variety of different
`chemistries.
`including DNA binding dyes. Roche
`Molecular Systems‘ TaqMan®. molecular beacons. dual
`FRET. and primer-based technologies, The optical design
`promotes the multiplexed analysis of up to four different
`fluorophores without compromising sensitivity or
`dynamic range. Bio-Rad also offers a personal-sized
`cycler. the Gene Cycler, which features a small footprint
`and a heated lid.
`
`Brinkmann Instruments: Westbury. MHZ-based
`Brinknrann‘s Eppcndoriw Mastercyclcrql line of thermal
`cyclers includes both personal-sized and gradicnl mod—
`els. All feature heated lids and are sold with a PCR
`license. The gradient model provides constant romping
`rates (SteadySlope1M technology} so that programs opti-
`mized on the instrument can be used on non—gradient
`machines.
`
`Cepheid: ACCording to company president Kurt
`Petersen. Sunnyvale. Calif.-bascd Cepheid has sold
`more than 400 Smart Cycler® instruments since the
`product was launched in May 2000. The system is based
`on the company‘s l—CORE® technology—truerorluidics—
`based. temperatureecontrolled modules that permit each
`sample to be subjected to different experimental condi~
`tions. Petersen explains that thc Smart Cycler is the only
`real-time system on the market that offers this "random
`access" capability, a feature that speeds up assay opti-
`mization by allowing researchers to operate each reaction
`site independently of the olhcrs. Each l-CORE unit can
`examine up to four targets in a reaction tube. thus facili-
`tating multiplex reactions.
`
`Cnrhett ResearCh: For gadget-loving researchers.
`Corbett Research of Sydney. Australia offers the Palm
`Cycler, an individual vsized thermal cycler integrated with
`a Palm PC. The instrument offers a gradient function fea-
`turing a true gradient from l-S degrees across the block.
`Corbett's Rotor—Gene, a
`real-time PCR system, can
`detect a variety of chcmistrics including SYBR Green 1
`[from Eugene. Orexbascd Molecular Probes). TaqMan.
`and molecular beacons. and can detect four different llu-
`crescent probes in a single tube. The Rotor-Gene
`maintains temperature homogeneity via a centrifuge
`rotor that spins the samples at 500 rpm.
`
`Idaho Technology Inc: This Salt Lake City based com-
`pany's latest offering is the Indy“l Air Thermocycler,
`which uses air cycling to decrease ramping and cooling
`time; a 30-cycle. three-temperature reaction can be come
`pictcd in lcss than 35 minutes. Unlike the company's
`flagship RapidCycler®. which uses a similar cycling
`technology but is limited to capillary tubes, the Indy ther-
`
`mocycler is designed for standard 24 it 0.2-rnl PCR
`tubes. According to company representative Nathaniel
`Lallalin. the Indy then'nocycler's speed and small foot-
`print make it
`ideal for research laboratories that are
`trying to increase throughput but do not wish to invest in
`a more expensive.
`large—capacity machine.
`Idaho
`Technology‘s popular RapidCyclcr model
`is still avail;
`able and is actually faster than the Indy cycler owing to
`the higher heat conductivity of glass capillary tubes;
`however. researchers may find that standard PCR tubes
`are easier to use.
`
`Marsh Bio Products: Rochestcr. N.‘r'.—bascd Marsh
`offers two Duncan thermal cyclers. each utilizing a water
`bath for thermal control. The DT—M and DT-IOS instru-
`ments can accommodate 24 or 108 microtilcr plates
`respectively. Since these instruments use water baths for
`temperature control. they exhibit small weil-to-wel] tem—
`perature variability. according to David Witkoski,
`Marsh’s marketing communications manager. The
`Duncan thermal cyclcrs are robotic—based and are mostly
`found in pharmaceutical and biotechnology companies.
`
`MatriCal: Chadds Ford. Pa.-bascd MalriCal developed
`the MatriCyclch“ for
`the company‘s specialized
`MarriFlaresW. which are designed to optimize the shape
`of high-throughput wells for increased signal detection
`and ease of liquid handling According to company liter-
`ature. researchers using the MatriCyclcr with the plates
`can increase their throughput up to |00 percent. with a
`reduction in reagent use of approximately 80 percent.
`Company co-t‘ounder Kevin Oldenburg explains that the
`MatriCycler overcomes four problems associated with
`the use of traditional cyclers for high~throughput experi-
`ments. First, the hearing block of a conventional cycler is
`a large thermal mass that takes a long time to hear and
`cool. Similarly. the plastic used in multi—wcll plates is
`thermally insulated and therefore takes time to heat.
`Because plastic tends to deform upon heating. the plates
`tend to warp and become wedged into the machine. pre-
`venting easy removal. Finally. reactions. using mulri-wel]
`plates require the use of a heated lid to prevent evapora-
`tion:
`this often results
`in the development of a
`temperature gradient across the plate.
`The MatriCyclcr works by healing each sample well
`with a thermally conductive probe. rather than relying on
`the thermal block to heat the samples. This eli minutes the
`problem of sample evaporation. speeds up sample heat-
`ing, and. according to Oldenburg. provides a heating rate
`of up In 20"C per second. Thus. a 30—cyclc reaction takes
`2040 minutes to complete with this instrument.
`
`MJ Research lrrc.: Ml Research lMlR) of Waltharn.
`Mass" has added a number of new items to the world of
`thermal cyclcrs. All of the company‘s cyclers feature
`Pelu'er heating and rttlphaTM modules.
`interchangeable
`
`Agilent Exhibit 1234
`
`Page 1 of 5
`
`Agilent Exhibit 1234
`Page 1 of 5
`
`
`
`healing blocks that allow users to change sample format
`rapidly. MIR offers [0 dillerent Alpha units for formats
`ranging from OAS-ml PCR tubes to microwell plates. to
`microscope slides; one of the more unique products is a
`set of microscope slide blocks that allow users to run a
`total of 32 slides at a time for in Situ PCR. The DNA
`Engine1M cycler. the company's “workhorse" model. fea—
`tures a heated lid and independent dual blocks to allow
`two experiments to be run simultaneously. The DyedTM
`thermal cycler. one of the newest members of the MIR
`family. allows four separate protocols to he run at the
`same time with the use oflwo Alpha modules. It also fea-
`tures an easy-to—use. Windows-driven, graphical user
`interface. The newer TetradTM model offers the same fea-
`tures as the smaller DNA EngineTM cycler. but is capable
`of handling four limes as many samples. All units feature
`a heated lid and are roboticcompatible. For more modest
`budgets, MJR offers a number of personal-sized cyclers,
`including the Mini‘Cycler and P'Ttllltlfl'l'M thermal
`cycler. These cyclers feature Hot BonnetTM heated lids
`and can be used for a variety of sample formats.
`MJR‘s recently released DNA Engine OpticonTM is a
`real-time cyclcr that uses fluorescent detection to moni~
`lor reactions. The system is essentially a DNA Engine
`chassis coupled to a 96-well Alpha unit and an optical
`detector. According to vice president of sales and mar-
`kcling Mike Mortiltaro. the Opticon is the ortly 96-well
`real time system on the market that involves no moving
`pans yet allows the instrument to view each sample or
`well independently. A photomultiplier tube detects sig-
`nals from samples
`indiVidualiy. "One of the big
`advantages that we see in the Opticon’s design is that
`we‘re looking at each sample independently and individ-
`ually. and We have much lower cross-talk. That gives us
`a broader dynamic range and a higher level of sensitivi-
`ty." says Mortillaro.
`
`MWG-Biotcch AG: Ebcrsburg. Gennany—bascd MWG.
`offers the Primus system. a multi-block cycler that
`is
`designed as a four‘hloek. standalone unit but can also be
`interfaced with up to 30 individual blocks via PC soft-
`ware. The. system also features a patented highrpressure
`lid that. unlike other heated lids. prevents evaporation of
`reaction volumes from SBd-wcll plates. Standard heated
`lids were developed to promote temperature homogeneity
`across the block. but were designed when scientists were
`still using individual microfuge tubes with caps. says
`Craig Abrahams. president of High Point. N.C.-bascd
`MWG Biotech luc..
`the company‘s U.S. subsidiary.
`However. in today‘s higher-throughput market where sci-
`entists use microplales. they cannot be fully sealed or
`capped easily. "So these same heated lids
`are now pro-
`moting evaporation." explains Abrahams. The Primus
`cycler’s highvpressure lid provides up to 300 Newtons of
`pressure. making it effective for evaporation protection.
`According to Abrahams. the Prirnus system has been pop-
`ular with high~throughput labs looking for high capacity
`instruments at a low cost. For example. Graham Casey.
`a
`researcher at
`the Cleveland Clinic Foundation in
`Cleveland. Ohio. recently purchased three of the multi-
`blocit instruments and says that he has been “pleased with
`all of the machines." Laura Bedzyk. staff biologist at
`Wilmington, Delxbased DuPont Co., has been using the
`Primus thermocycler in her research for almost two ycurs
`for generating PCR products for DNA microanays;
`Bedzyk says that the factors influencing her decision to
`purchase this insu‘ument included “multiple. independent
`positions. pressurized lids. ease of use. and the potential
`for incorporation into a custom. automated system."
`
`Indianapolis—based
`Roche Molecular Biochemicals:
`Roche has licensed Idaho Technology‘s thermal cycler
`technology and reengincered it
`for the LightCyclerTM
`insu-ument. a real-time thermal cycler that. like Idaho‘s
`RapidCycler. employs circulating air and glass capillary
`tubes to dramatically reduce amplification time. The sys-
`tem uses fluorescent detection for real-time analysis and
`is controlled by a PC workstation. Researchers can use
`the LightCycler for assays using SYBR Green I.
`
`LabConsumer
`
`
`
`Roche‘s Ughtfycl’er instrument uses circulating air and
`glass capillary tubes to reduce amplification time.
`
`sequence-specific hybridization probe pairs. or several
`other detection formats.2
`
`St. John Associates: Beltsvitle. Md-based St. John's
`BioDven lll Therrnocycler is based on circulating air
`technology but is capable ofhandling samples in a vari-
`ety of formats,
`including standard PCR tubes. glass
`capillary tubes. slides. and multiwell plates. Designed
`for very high capacity. the BioOvcn Ill cycler can han—
`dle ll) multiewell plates per run and features a rotating
`carousel lo ensure lempcrature uniformity. According lo
`Peter St. John. the BioOven liI cycler enjoys populari-
`ty with scientists in a variety of
`fields including
`agribusiness.
`forensics. and pharmaceutical
`research.
`because it offers greater well-to-well uniformity than
`aluminum block cyclcrs and provides higher flexibility
`in terms of sample media.
`
`.lnllu. Calif-based Slratagene's
`Stratagenc: La
`RohoCycler
`features
`four programmable blocks and
`offers a gradient feature to simplify optimization What
`makes this cycler unique is that it employs a robotic arm
`to move samples from block to block. The user can set
`three of the four blocks to denaturation. annealing. and
`polymerization temperatures; the fourth is a cooled stor—
`age block. Because samples are moved rn‘notically to the
`appropriate temperature block for each point in the reac-
`tion. there is no need for temperature ramping of a given
`block. According to company literature this method
`reduces experiment time up to 3G perccnt. Product man—
`ager Peter Pingerelli explains that the popularity of the
`RoboCycler stems from its durability. Competing instrue
`merits. he says. “have had a lot of problems. with
`heavy-use instruments [failing] post-warranty."
`Stratagcne's deUUOT” Multiplex Quantitative PCR
`System, introduced last year:2 offers fluorescence-based
`real-time detection and can measure up to four different
`dyes in a single tube. The system uses a quartz-tungsten
`halogen lamp rather than a laser light source. broadening
`the excitation wavelength range.
`it also features a PC
`interface and. according to company literature. tempera-
`ture control that exceeds the industry standard,
`
`Techne: Princeton. N.J.-based Techne offers a range of
`cyclcrs to fit different research needs and budgets. The
`Techgene'E cycler. a Peltier-bascd system Ihat replaces
`the company‘s Progene® model. is a compact. inexpene
`stve instrument that researchers can run remotely through
`a PC Although the Techgene cycler is designed for a low
`sample quantity (2&35 wells). up to 32 units can be
`linked for larger experiments. The Flcxigenefl" cycler.
`which will replace the company‘s Genius® model.
`is a
`Peltier—based system designed for higher throughput.
`
`Technc‘s Touchgcne® Gradient Thermal Cyclcr is adver-
`tised as the most linear gradient cycler on the market—it
`is linear over a 30°C range —and offers real-time graphi~
`cal display and a touch—sensitive screen for ease of
`programming. To
`ensure
`block
`uniformity,
`the
`Touchgene cycler includes four temperature sensors per
`block. All Techne eyelets come standard with adjustable
`heated lids and interchangeable blocks.
`
`Therme Hybaid: Franklin. Mass-based Thermu
`Hyhaid‘s cyclers, which range from individualvsized to
`multi—block networks. come in a variety of formats.
`The company‘s most basic system, the PCR Sprint. fea
`tures Peltier heating. a 20-24 sample individual block,
`and two different block format options. Thermn
`Hybaid‘s PCR Express and soon-lo-be—relcased Pall.
`also Pelticr-bascd machines. feature six different inter-
`changeable block options. including a flat block for in
`situ or microaIray experiments. The lel also includes
`Thermo Hybaid‘s new last block technology and a
`large graphical display panel.
`The company‘s MultiBloel-t System (MES) is a flexi-
`ble thermal cycler system driven and monitored by a
`central PC that features software. which allows real-time
`monitoring and automatic logging for good laboratory
`practice (GLPl purposes. By enabling up to 30 individual
`blocks to be attached to a single processor. users pay less
`per block than they would if purchasing individual stand-
`alone machines And. unlike the multi-bloclt systems
`from MJR and MWG, Thermo Hybaid‘s system allows
`the user to purchase blocks individually. The company's
`newest product
`is
`the MES“.
`the
`fully robotic
`MultiBlock. which facilitates high-throughput PCR.
`According to the company. by ejecting the whole block
`through the front of the unit, the M33“ is fully slackable
`and integrates easily into robotic systems. E
`Aileen Carma”: can be contacted at
`at'ortsmnslé‘the-scierrristrom.
`References
`I. G. Metsenholder. "New lids I.ll'l the block.“ The Sch-nun.
`Nov. 22. 1999
`2 D. Fitzgerald. "Up to speed on PCR." The Scientist. 14[2]i'31. Nov
`27'. 20ml.
`3. A Constans. ”Court: Last clouds met P'CR pricing." The St'tellftll'.
`Iiitii:l.st-pt 3.20m
`
`|3I33]:l7.
`
`
`
`St. John ‘5 BioOven ll'l' thermoqcler offers
`flexibility in terms ofsample fem-tats.
`
`The Scientist 33
`December in. 200]
`
`Agilent Exhibit 1234
`
`Page 2 of 5
`
`Agilent Exhibit 1234
`Page 2 of 5
`
`
`
`LabConsumer
`
`Suppliers of Thermal Cyclers _
`
`Applied Biosystems
`{800) 345-5224
`www‘appliedbiosystemsmm
`
`Biometra
`my:
`>I.5/>L3
`+ 49 [551) 50 68 6 0
`www.biometra.wm
`4.5} 3.0
`+.-'- 0.31
`
`
`+i-033f
`
`4.1! 3.0
`
`3I|i3I|
`
`[amhi‘hrmlc
`
`BiorRad
`(BUD) 41251-5723
`\mm.hio-rad.com
`
`Brinkmann
`[800) 645-3050
`wmvlninkmann‘com
`
`M {M
`— 7 l—I
`Mic“
`mm
`
`10f”
`Pehiu
`
`
`Pelvis!
`
`fallier
`
`1|]! HI
`
`31]f1.0
`
`
`
`
`
`
`vmijdahotechxom
`
`Cepheid
`(333,1 538 3222
`wuwu.smartcyclermm
`Corbett Research
`+61(2)9?36132|]
`www(urbettresearduom
`
`Uniofipmtm'nthutks
`
`
`W
`
`Famine hauler mlh 1m
`mlmg
`
`
`6L Appfled Research
`(IS-'17) 22372220
`
`Mug
`WW I“
`
`- ,
`EK-Hla’auflcflmndtfir)
`
`mum
`
`
`Idaho Tec|1nomgy
`(300) 735-65413;
`
`.
`Wilhelm
`
`
`34 The Scientist
`December [0. 200i
`
`Agilent Exhibit 1234
`
`Page 3 of 5
`
`Agilent Exhibit 1234
`Page 3 of 5
`
`
`
`LabConsumer
`
`Suppliers of Thermal Cyclers (continued)
`
`Marsh Bio Produnr.
`{800} 5-2812
`www marshbiucum
`
`.
`
`-
`
`,
`
`,
`
`
`
`
`
`
`
`
`MatIiCdl
`(E10136
`www.mat
`MJ Research
`(333] PELTIER
`www.mj ncom
`
`. ' '
`
`MWG BiEJtL-L'h
`(37?) 69
`33?.
`www.ruwgblulenlmcmn
`Rnche Mn‘erulm‘ Binilwmicals
`{300) 262-1
`|Jiochem.roche_cum
`St, John Assocmtes
`{80
`Fifi-2
`‘
`www.suohnassociatesxorn
`Straugene
`-
`‘
`.
`.
`{800142 -5
`at
`w.rw.5tra ._ genecom
`
`lawn»
`
`fllllE-filhnflph
`
`1.01111
`
`:1“ ELM
`
`S—Iflfl
`
`.
`
`.
`
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`
`
`
`
`
`mum
`
`u‘ 03°: WNW
`
`.
`
`MI3his'
`I
`W I'
`”(mf’d' mum ”a";
`”
`
`Mydu 3mm 1mm
`mum“
`
`an» 1-3; Resmive
`m 4; Mn" mm
`
`Lolm
`
`sow-mm
`
`u.
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`'
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`
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`v-l- 05°:
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`Tecime
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`
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`Thermo Hybaid
`(3881 JVHYBAID
`umrwlhermohyba
`
`Editors Note: To the best of our knowledge. this tabfe is a comprehensive fistr'ng of suppliers.
`
`
`The Scientist 35
`December 10, 2001
`
`Agilent Exhibit 1234
`
`Page 4 of 5
`
`Agilent Exhibit 1234
`Page 4 of 5
`
`
`
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`Agilent Exhibit 1234
`Page 5 of 5
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