Covaris ML230 Manual

ML230 Focused-ultrasonicator
Setup & Instrucon Manual
Instrument Developed for High-performance
Sample Preparaon using Adapve Focused Acouscs® (AFA®)
For Research Only
Model Number: ML230 | Soware: SonoLab 10.1
Covaris
Part Number: 010516 Rev B | March 2020
ML230 Setup & Instrucon Manual

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Part Number: 010516 Rev B | March 2020
ML230 Setup & Instrucon Manual
UNIVERSAL PRECAUTIONS
Universal Precauons should be followed on all specimen samples, regardless of whether a sample is known to contain
an infecous agent. Laboratories handling specimen samples are advised to comply with applicable parts of the following
government and clinical standards, or their equivalent in the country of use:
• Centers for Disease Control (CDC), Universal Precauons for Prevenon of Transmission of HIV and Other Bloodborne
Infecons, published 1987, updated 1996
• Clinical and Laboratory Standards Instute (CLSI), GP17-A2 Clinical Laboratory Safety; Approved Guideline - Second
Edion, published 2004, ISBN 1-56238-530-5
• Clinical and Laboratory Standards Instute (CLSI), M29-A3 Protecon of Laboratory Workers from Occupaonally
Acquired Infecons; Approved Guideline, Third Edion, published 2005, ISBN 1-56238-5674
• Occupaonal Safety and Health Administraon (OSHA), 29 CFR 1910.1030 Bloodborne Pathogens
• Internaonal Standards Organizaon (ISO) 15190:2003, Medical Laboratories – Requirements for Safety
Trademarks
Patented or patent pending and registered or registraon-pending trademarks of Covaris are protected.
Registered names and trademarks used in this document, even when not specically marked as such, are not to be considered
unprotected by law.
WCS and AFA are either registered trademarks or trademarks of Covaris, Inc. in the United States and/or other countries.
Teon is a registered trademark of The DuPont Company.
Informaon subject to change without noce
For Research Use Only (RUO)
Not for use in Diagnosc Procedures
Publicaon PN 010516
Product covered by Patent No. US6, 719,449, US6, 948,843, and other pending applicaons
2020© Covaris Inc., All Rights Reserved
Covaris, Inc.
14 Gill Street, Unit H
Woburn, Massachuses
01801-1721 USA

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ML230 Setup & Instrucon Manual
Warnings
For the Safety of Operang Personnel:
• Make sure that the equipment is properly grounded. DO NOT operate if it is not properly grounded.
• The unit is equipped with a power plug appropriate for the desnaon country. DO NOT, under any circumstances, remove the
grounding prong from the power cord.
• DO NOT aempt to operate the equipment with the safety cover in the open posion or without water in the acousc
assembly; the acousc system will not work. If there is any indicaon that the Safety System is not funconing properly,
DO NOT operate the equipment and contact Covaris immediately.
• If the equipment is used in a manner not specied by the manufacturer, the protecon provided by the equipment may be
impaired.
To Prevent Damage to the Equipment:
• The instruments are designed to operate in ambient laboratory condions e.g., 19 ºC to 25 ºC (66 ºF to 77 ºF).
DO NOT operate the instrument in a cold room environment; the system is designed to operate with a water bath and includes a
solid state heater/chiller apparatus to control sample temperature.
• NEVER run a method without water in the acousc assembly; this could damage the transducer. The instrument is equipped
with a water level sensor to protect the transducer. The system will not allow the acousc wave treatment to start unless an
adequate volume of water is detected.
• DO NOT leave water in the water bath with instrument power turned o for an extended me as this disables the water
cleaning system within the apparatus.
• AFA-grade water should be used to ll the supply water bole.
• DO NOT employ isopropyl alcohol, ammonia-based or abrasive cleaners on the acousc assembly.
• Sample plates should be stored in a dry place when not in use.
• DO NOT load third party hardware, soware, or parts onto the system without consulng with Covaris.

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ML230 Setup & Instrucon Manual
Warranty
When used in accordance with wrien instrucon and under normal operang condions, the Covaris instruments are guaranteed
to be free of defects in MATERIAL and WORKMANSHIP for one (1) year from the date of original delivery by an authorized
representave. Any component proven defecve during the stated period will be repaired free of charge or replaced at the sole
discreon of Covaris, F.O.B., Woburn, Massachuses, U.S.A. provided that the defecve component is returned properly packaged with
all transportaon charges prepaid. The customer is expected to perform basic diagnoscs and component replacement via telephone
support from Covaris personnel. If Covaris personnel are required to perform on-site repair, all travel related costs are paid by the
customer. A limited warranty as specied may apply to certain components of the equipment.
Warranty Excepons
This warranty is void if failure of the soware or hardware has resulted from accidents, abuse, improper maintenance or repair, or by
misapplicaon from the customer. It is also void if damage is caused by any unauthorized aachments or if modicaons are made to
the equipment. Removing or tampering with the Safety Enclosure will void the warranty, and the customer will assume all liabilies.
This warranty is limited to the original purchaser and is not transferable.
The soware will perform according to the accompanying wrien materials and the medium on which the soware is delivered is
free of defects in materials under normal use and service. The warranty is void if damage has resulted from third party soware not
intended for use with the system.
The high-power focused transducer is designed to give maximal mechanical energy output in water. Permanent damage to the
transducer and electronic circuits could result if the transducer is operated without water. Operaon of the system without water in
the water bath voids the warranty.
CONTACT COVARIS, INC. SHOULD YOU HAVE QUESTIONS OR CONCERNS PERTAINING TO THE USE OF OUR EQUIPMENT.
Warranty Services
The purchased equipment is covered by a twelve (12) month warranty which includes all the necessary services and support necessary
so that the customer can successfully operate their equipment. Extended warranes are available at the end of the original 12 month
warranty period.
Services included with the original purchase of the system are:
Technical Support – Ongoing assistance with the operaon or applicaon of the equipment and/or troubleshoong is provided via:
• Telephone
- United States: Tel: [phone] during the hours of 9:00 a.m. to 5:00 p.m., Monday through Friday,
Eastern Standard Time (EST), Greenwich Mean Time (GMT-05:00)
- Europe: Tel: [phone], during the hours of 9:00 a.m. to 5:00 p.m., Monday through Friday, Greenwich Mean Time
• E-mail queries to [email protected] or applica[e-mail]
ML230 Instrument Repair – Warranty repair of the ML230 is provided on site. All labor and expense charges are included. Failure due
to accident, abuse, or improper operaon is not covered. An Operator’s Manual is provided with the equipment. This manual includes
secons on the operang instrucons, maintenance guidelines, and troubleshoong ps.

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Table of Contents
1. Introducon ...................................................................................................................................................................................... 6
1.1 Overview of the Instrucon Manual ................................................................................................................................. 6
1.2 The ML230 and the Covaris Process ................................................................................................................................ 6
2. Installaon and Setup ..................................................................................................................................................................... 7
2.1 Unpacking the System ......................................................................................................................................................... 7
2.2 Assembly ................................................................................................................................................................................ 7
2.3 Levelling the ML230 Instrument ........................................................................................................................................12
2.4 ML230 Soware ................................................................................................................................................................... 13
2.5 Starng the ML230 Instrument ......................................................................................................................................... 14
3. Operaon .......................................................................................................................................................................................... 15
3.1 General Overview ................................................................................................................................................................. 15
3.2 Preparing the ML230 for Use ............................................................................................................................................. 16
3.3 ML230 Water Control .......................................................................................................................................................... 19
3.4 Loading the Sample .............................................................................................................................................................. 19
3.5 Running a Method ................................................................................................................................................................ 21
3.6 Instrument Light .................................................................................................................................................................... 21
3.7 User Maintenance ................................................................................................................................................................. 22
3.8 Shutdown Procedure ............................................................................................................................................................ 23
4. Using a Pre-installed Method ........................................................................................................................................................ 24
4.1 DNA Shearing Protocols ...................................................................................................................................................... 24
5. Create and Edit Methods ............................................................................................................................................................... 25
5.1 Create New Method ............................................................................................................................................................. 25
5.2 Modify Exisng Method ...................................................................................................................................................... 30
6. SonoLab™ Applicaon Soware .................................................................................................................................................... 32
6.1 Method Tab ............................................................................................................................................................................ 32
6.2 Schedule Tab .......................................................................................................................................................................... 34
6.3 History Tab ............................................................................................................................................................................. 36
6.4 About Tab ............................................................................................................................................................................... 37
6.5 Admin Tab ............................................................................................................................................................................... 38
7. System Specicaons ...................................................................................................................................................................... 42
8. Troubleshoong ............................................................................................................................................................................... 43
9. Appendix ........................................................................................................................................................................................... 44
Appendix A: M-Series Site or Transport Preparaon .............................................................................................................44
Appendix B: Updang SonoLab Soware ............................................................................................................................... 44
Appendix C: Replacing the ML230 Parcle Filter Element (PN 500679) ..........................................................................45

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ML230 Setup & Instrucon Manual
1.1 Overview of the User Manual
This manual contains the installaon, operaon, and service instrucons for the ML230 Focused-ultrasonicator which are essenal
to the proper use and care of this equipment. Should any unforeseen problems occur with the normal operaon of the equipment,
contact Covaris Technical Support ([e-mail]) immediately.
The following denions apply in this manual:
1.2 The ML230 and the Covaris Process
The ML230 Focused-ultrasonicator is based on Covaris Adapve Focused Acouscs (AFA) technology. This instrument is designed to
provide researchers an integrated stand-alone tool to be used in laboratory environment. The instrument is ideal for those who work
with DNA shearing and sample preparaon.
The system is comprised of the Treatment System which delivers the acousc energy to the sample, the Safety System which protects
users from inadvertently contacng the high intensity acousc energy, and an embedded Computer and Applicaon Soware which
provides users an interface to control the device.
The Covaris Process uses Adapve Focused Acousc energy to precisely control cavitaon and acousc streaming within the sample
treatment vessel in a non-contact, isothermal fashion. Focused acousc energy is generated and delivered to individual samples. The
mechanical energy imparted on the sample results in a controlled series of compression and rarefacon events.
1. Introducon
NOTE: Informaonal. CAUTION: Equipment damage may occur.WARNING: Personal injury may occur.

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2.1 Unpacking the System
Follow the included instrucons for removing the ML230 instrument and accessories from their shipping materials.
2.2 Assembly
The following items should be found in the package of the Covaris ML230 instrument:
• ML230 main unit
• Accessory Box
- Bubble Level Tool
- Wrench for removing the caps/vents from the water boles
- Sample Tray Assembly
- 2 Plasc water boles
- 1 ML230 power cord
- Sample Racks and Consumables, depending on order conguraon
2. Installaon and Setup
Figure 2.1 Leveling Tool and Reservoir Cap Wrench
Figure 2.2 Sample Tray Assembly
CAUTION: The ML230 weighs about 80 pounds (36 kg). With the front of the instrument closest to your body, li the
ML230 by gripping from its base plate. Liing the unit by its plasc case or doors will damage the instrument.

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Aer the instrument is unpacked, posion the ML230 on a at surface near a power outlet. Use both sides of the base to li the
instrument. DO NOT li by the plasc parts at the front of the instrument.
1. Assemble the Water Control supply and waste boles.
• Locate the two plasc boles and the bole cap tool in the accessory box.
Figure 2.3 Front view of the ML230 instrument
Figure 2.4 Water Control System Bole
Touch Screen
Control Panel
Safety Cover
Leveling Feet
Resevoir and Drain
Water Boles
Sample Handling
System behind
Safety Cover
Vented Filling Cap
toward rear of ML230
Quick Disconnect
Fing towards
front of ML230
Accessory Drawer

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• Remove the vented cap on the ‘Supply’ bole using the cap wrench and ll the bole with AFA-grade water. Replace the
cap using the cap wrench. DO NOT over-ghten the cap.
• Holding the two quick-connect tubes up and out of the way, slide the empty Waste bole into the le-most opening at
the front of the ML230. The vented cap is toward the rear; the Quick Disconnect cap toward the front of the instrument.
• Slide the Supply bole lled with water into the remaining opening, between the Waste bole and the sample
handling area.
Figure 2.5 Using Cap Wrench to Secure Cap
Figure 2.6 Waste drain and ll port
NOTE: The caps for ‘Waste’ and ‘Supply’ reservoirs are not interchangeable. The waste cap is marked ‘Waste’ and
contains a relief valve as shown below. There is no relief valve in the ‘Supply’ cap.
No Valve in Supply Cap
Waste Cap Contains a
Relief Valve

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• Press the two Quick Disconnect ngs into the Waste and Supply boles unl they click into place. See Figure 2.4.
2. Assemble the Sample Tray Assembly to the ML230.
• Locate the Sample Tray Assembly in the accessory box
Figure 2.7 Boles into Water Works Bay
Figure 2.8 Connecng Water Works Bole to System
Figure 2.9 Mounng Bracket
Press ng on ML into
mang ng on bole
unl it clicks into place.
Holes for Locang Pins
Located on Rear Face
of Mounng Bracket

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• Li the Safety Cover open.
• Slide the Sample Tray Assembly locang pin holes onto the pins on the moon system, behind the safety cover.
The sample tray faces to the le. Refer to the image below.
Figure 2.10 Mounng Bracket
Figure 2.11 Mounng Bracket installed in ML230
Locang Pin Holes
Locang Pins
Mounng Bolt

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• Thread the Sample Tray Assembly’s thumbscrew into the moon system and snug it nger-ght.
3. As shown in Figure 2.13, plug the AC power cord into the power inlet located about half-way up the le side of the ML230 rear
panel. Plug the AC power cord into the power outlet.
The enre instrument may be relocated in this assembled conguraon by disconnecng power, liing from the boom, relocang, and
repeang the appropriate assembly steps.
2.3 Leveling the ML230 Instrument
This step is very important to ensure proper lling and draining of the water bath and proper instrument operaon. Once the ML230
is installed in the locaon in which it will be used:
• Open the Front plasc safety cover
• Place the leveling xture in the sample tray
Figure 2.12 Sample Tray Assembled to ML230 Figure 2.13 The power panel of the
ML230 instrument
NOTE: Any me the ML230 is moved to a new locaon, follow the instrument leveling procedure described in Secon 2.3
Sample Tray Assembled to
ML230 Moon System
Snug Thumb Screw
to Finger-ght
Power Inlet for
AC Power Cord
Power Switch
Figure 2.14 Leveling Tool

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2.4 ML230 Soware
SonoLab 10 has been pre-installed on the ML230 instrument. No further installaon or conguraon is required.
SonoLab is the soware used to control a Covaris Adapve Focused Acouscs (AFA) instrument for processing a sample. It is designed
to create, edit, and run a sample processing program called a method. It has provisions to run, pause or stop a method and view
execuon histories which contain a record of key instrument parameter values during a run.
If it is necessary to install or re-install SonoLab, please contact Covaris for the latest versions, downloading, and installaon
instrucons.
• If the bubble is to the right side of the center markings of the sight glass, the right side is high, and the right side feet need to be
shortened. If the bubble is to the le side, the right side is low, and the right side feet must be lengthened. Adjust both front
and rear feet an equal number of turns.
• Repeat the process above to level front-to-rear, adjusng only the feet in the front. If the bubble is in the center of the xture
sight glass, no adjustment is required. If the bubble is to the front side of the center markings of the sight glass, the front
side is high, and the feet need to be shortened. Alternavely, if the bubble is to the rear, the front side is low, and the legs must
be lengthened. Adjust both front feet an equal number of turns. The feet can be adjusted by hand; no tools are necessary.
• Lengthen or shorten both legs on the right side of the instrument an equal amount, as in Figure 2.15, unl the bubble is
centered in the leveling xture sight glass.
Figure 2.15 ML230 Leg Height Adjustment
NOTE: The feet can be adjusted by hand; no tools are necessary.
Turn both legs
clockwise to shorten
Turn both legs counter
clockwise to lengthen

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2.5 Starng the ML230
Turn on the ML230 power switch located about half-way up the le side of the rear panel.
SonoLab will automacally connect to the instrument, and a window will appear asking whether the instrument is to be homed. Touch
“Yes”. The moon system will perform its inializaon sequence, the homing window will close, and the SonoLab Home screen will be
displayed, as illustrated in Figure 2.16.
Figure 2.16 The SonoLab Method Screen
NOTE: The ML230 is shipped with the sample tray removed. If the sample tray has not already been installed, follow the
instrucons from Secon 2: Installaon and Setup.

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3.1 General Overview
The Safety Cover is used to protect users from potenal acousc hazards during the treatment. The acousc treatment will not be
started if the Safety Cover is open.
The Acousc Assembly, located in the water bath below the sample tray, is used to transmit the acousc energy from transducer
to the sample through the coupling media (AFA-grade water). The assembly incorporates an Acousc Transducer which converts
electrical signals into ultrasonic acousc signals. The assembly also incorporates Sensors for water level and temperature to ensure
that these are appropriate during acousc treatments.
The 2-axis Moon System is designed to automacally align the sample tube at the appropriate posion and height in the acousc
eld for processing. Tubes are loaded into appropriate Racks, which are then placed on the ML230 sample tray.
An Embedded Processor provides an interface for controlling the ML230 instrument via the touch screen at the front of the ML230.
The Electronics Cabinet provides the hardware for controlling the acousc transducer, the temperature, and sample posion during
processing.
The Enclosure Light is normally lit when the instrument is turned on. The user can disable the light from the SonoLab Home screen.
3. Operaon
Figure 3.1 Principal components of the ML230 instrument
Touch Screen
Control Panel
Safety Cover
Leveling Feet
Waste Bole
Supply Bole
Sample Handling
System behind
Safety Cover
Accessory Drawer

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The Water Control consists of a pump, valves, supply and waste boles, and interconnecng plumbing. These components are
controlled automacally by SonoLab soware to ll, drain, and maintain the correct water level in the water bath.
When the bath is lled, the water is circulated through a UV sterilizer and parculate lter to prevent biological growth in the water
control hardware.
The Thermal Module controls the water bath temperature. It cools or heats the acousc assembly, maintaining the water bath at or
near the setpoint programmed in the method. No external chiller is required.
3.2 Preparing the ML230 for Use
To run an acousc treatment, a number of condions must be met, including:
a. The ML230 moon system must be inialized
b. There must be an acousc method selected in the Method box
c. The proper rack and consumables must be in place on the ML230 sample tray. See Secon 3.5, Loading the Samples
d. Water Temperature must be within acceptable Min and Max limits
e. Water must be at the level required by the consumables specied in the selected method
f. The safety cover must be closed
The Run buon will remain gray and inacve unl all necessary condions except water level and temperature are met. The ML230
automacally corrects these when “Run” is selected.
To achieve an acceptable status on the other condions:
a. The ML230 moon system normally inializes when SonoLab starts, unless the user purposely skipped this step, in which case
the moon system can be homed from the Moon Control pane.
• Touch the Moon Control buon to display the Moon Control pane
• Touch “Home” to inialize the moon system
WARNING: During treatments, objects immersed in the focal zone above the transducer will be exposed to high acousc
intensity when the system is acvated. Always close the Safety Cover prior to starng a method. Do not tamper with
the door-closed sensor system.
Figure 3.2 Moon Control Panel
Touch the Moon Control buon then touch Home

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b. To select and load an acousc method, touch the “Method” bar, as shown in Figure 3.3, to display the Method Select pane.
Navigate through the list with the vercal scroll bar, if necessary, and select a method.
The method’s total and remaining run me (minutes and seconds), temperature setpoint, and actual bath temperature will be
displayed on the main screen as shown.
c. The ML230 system automacally adjusts the water temperature to match the method temperature setpoint aer Run is
selected. Water Temperature status will turn green when temperature is within the Min and Max limits (typically 3 degrees
above or below the setpoint). The default setpoint (in the absence of a selected method) is 20 ºC.
d. The ML230 Water Control automacally lls the bath to the appropriate level for the tubes specied in the method when the
Run buon is touched to start a method. (See Secon 3.3)
NOTE: Several acousc methods will come pre-installed onto your system.
Figure 3.3 Selecng and loading an acousc method
Figure 3.4 Method Run Time and Setpoint
Touch to display Method Select pane
Treatment Time Remaining/
Total Treatment Time
Bath temperature and setpoint

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ML230 Setup & Instrucon Manual
e. The Cover Closed status will turn green as soon as the Safety Cover is completely closed.
The remaining condions, along with their related states, are displayed in the Instrument Status panel (see Figure 3.5). Touch the
“System Status” buon to display the Instrument Status panel. An acceptable condion is indicated by a green check mark (ü) in the
Status box. If a condion is not acceptable, the status will show (X).
If any of the items in the Status list display (X), a red X will also be displayed in the System Status buon, as shown in Figure 3.6.
Once a method has been loaded and all condions except temperature and water level are acceptable, the Run buon will turn green
and the selected acousc Method can be started.
To modify an exisng method, see Create and Edit Methods in Secon 5.
Figure 3.5 Instrument Status Panel – All OK
Figure 3.6 Instrument Status Panel – Moon System not Homed
A red X in any item(s) in the Status list
results in a red X in the System Status buon

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3.3 ML230 Water Control
To transmit acousc energy to the sample vessel, the bath surrounding the transducer must be lled with deionized water. Covaris
recommends AFA-grade Water. You can also use highly puried water (at least ASTM Type III or ISO grade 3).
The ML230 Water Control automacally maintains the correct water level in the bath for the sample vessels specied by a method.
This system either adds water from the Water Works supply bole to raise the level of the bath or removes water from the bath to the
Water Control waste bole.
Sensors detect if the supply bole is empty or the waste bole is full, and these condions are reported in the Instrument Status
panel. To service the boles, press the metal thumb latch on the Quick Disconnect Cap, as shown in Figure 3.7, to release the Quick
Disconnect Insert of the bole being serviced, and slide the bole out. Use the Water Bole Cap tool to remove the vented cap at
the rear of the bole, then empty the waste bole or ll the supply bole, as appropriate, and reverse steps to reinstall the bole. The
Supply bole is to the right, next to ML230 sample handling system, and the Waste bole is to the le of the Supply bole.
3.4 Loading the Samples
A variety of tubes and their associated racks are available for use with the ML230. In some cases, individual tubes are simply placed
into the rack as shown.
The 8 microTUBE strip is ed to its rack as a unit. Open the hinged rack and insert the 8 microTUBE strip. The strip is keyed to t
the rack in the correct orientaon.
Figure 3.7 Releasing the Quick Disconnect Fings
Press Metal thumb latch to Release
Quick Disconnect Insert
Metal thumb latch
Figure 3.8 Assembling microTube 8-strip into Rack
Tab on Strip and
Mang Notch on Rack

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Covaris
Part Number: 010516 Rev B | March 2020
ML230 Setup & Instrucon Manual
Close the hinged rack cover to secure the strip. Magnets in the cover hold the cover in the closed posion.
Open the ML230 safety cover. Place tubes and rack assembly into the ML230 sample tray shown in Figure 3.10. The racks are keyed
to load into the sample tray in the correct orientaon only.
Close the Safety Cover to ensure that the Door status in SonoLab turns to a green check.
Figure 3.9 microTUBE loaded in ML230 Sample Rack
Figure 3.10 Sample Rack Loaded onto ML230 Sample Tray