S1000 Thermal Cycler Instruction Manual Rev D

S1000 Thermal Cycler Manual

Copyright ©2009 Bio-Rad Laboratories, Inc. Reproduction in any form, either print or electronic, is prohibited without written permission of Bio-Rad Laboratories, Inc. Windows XP and Windows Vista are trademarks of Microsoft Corporation.

LICENSE NOTICE TO PURCHASER This base unit, Serial No. ____________, in combination with its immediately attached Bio-Rad sample block module(s), constitutes a thermal cycler whose purchase conveys a limited non-transferable immunity from suit for the purchaser’s own internal research and development and for use in applied fields under U.S. Patent No. 5,475,610 (claims 1, 44, 158, 160-163 and 167 only) or corresponding claims in their non-U.S. counterparts, owned by Applera Corporation. No right is conveyed expressly, by implication, or by estoppel under any other patent claim, such as claims to apparatus, reagents, kits, or methods such as 5’ nuclease methods. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.

Bio-Rad’s thermal cyclers and real-time thermal cyclers are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Nos. 6,767,512 and 7,074,367.

Hard-Shell PCR plates are covered by one or more of the following U.S. patents or their foreign counterparts owned by Eppendorf AG: U.S. Patent Nos. 7,347,977, 6,340,589, 6,528,302.

i

S1000 Thermal Cycler Manual

Bio-Rad Laboratories Resources Table 1 lists Bio-Rad resources and how to locate what you need. Table 1. Bio-Rad resources Resource

How to Contact

Local Bio-Rad Laboratories representatives

Find local information and contacts on the Bio-Rad Laboratories website by selecting your country on the home page (www.bio-rad.com). Find the nearest international office listed on the back of this manual

Technical notes and literature

Find technical information on the Bio-Rad Laboratories website (www.bio-rad.com). Type a term in the Search box and select Literature to find links to technical notes, manuals, and other literature

Technical specialists

Bio-Rad Laboratories provides quality technical support. We staff our Technical Support department with experienced scientists to provide our customers with practical and expert solutions To find local technical support on the phone, contact your nearest Bio-Rad Laboratories office. For technical support in the United States and Canada, call 1-800-424-6723 (toll-free phone) and select the technical support option

Warranty The S1000 thermal cycler and associated accessories are covered by a standard Bio-Rad warranty. Contact your local Bio-Rad Laboratories office for details of the warranty.

Safety and Regulatory Compliance The S1000 thermal cycler heats and cools very quickly during operation. We strongly recommend that you follow the safety specifications listed in this section and throughout this manual. Use only Bio-Rad USB cable (catalog #184-8000) when using any 1000-series cycler.

ii

S1000 Thermal Cycler Manual

Safety Warning Labels Warning labels posted on the instrument and in this manual warn you about sources of injury or harm. Refer to Table 2 to review the meaning of each safety warning label. Table 2. Instrument safety warning labels Icon

Meaning CAUTION: Risk of danger! This symbol identifies components that pose a risk of personal injury or damage to the instrument if improperly handled. Wherever this symbol appears, consult the manual for further information before proceeding CAUTION: Risk of electrical shock! This symbol identifies components that pose a risk of electrical shock if improperly handled CAUTION: Hot surface! This symbol identifies components that pose a risk of personal injury due to excessive heat if improperly handled

Instrument Safety Warnings The following warning labels display on the instrument and refer directly to the safe use of this S1000 thermal cycler (Table 3). Table 3. Instrument safety warning labels Icon

Meaning Warning about risk of harm to body or equipment. Operating the S1000 thermal cycler before reading this manual can constitute a personal injury hazard. Only qualified laboratory personnel should operate this instrument Warning about risk of harm to body or equipment from electrical shock. Do not attempt to repair or remove the outer case of this thermal cycler base, power supply, heat pump, or other accessories. If you open these instruments, you put yourself at risk for electrical shock and void your warranty. All repairs must be done by an authorized repair service Never remove the outer case of a thermal cycler base. This may cause you to receive an electrical shock. This thermal cycler uses neutral fusing, which means that live power could still be exposed inside the instrument even when the fuse is blown or removed Warning about risk of burning. A thermal cycler generates enough heat to cause serious burns. Wear safety goggles or other eye protection at all times during operation. Always allow the sample block to return to idle temperature before opening the lid and removing samples. Always allow maximum clearance to avoid accidental skin burns Warning about risk of explosion. The sample blocks can become hot enough during the course of normal operation to cause liquids to boil and explode

iii

S1000 Thermal Cycler Manual

Safety and Regulatory Compliance This instrument has been tested and found to be in compliance with all applicable requirements of the following safety and electromagnetic standards (Table 4). Table 4. Conditions for safe use Usage Aspect

Conditions for Safe Use

Rated input power

100–240 VAC, 50–60 Hz

Fuses

10 A, 250 V, 5 x 20 mm, fast blow (qty. 2)

Environment

Indoor use only

Temperature

15-31oC

Relative humidity

Up to 80% (noncondensing)

Altitude

Up to 2,000 meters above sea level

Overvoltage category

II

Pollution degree

2

SAFETY COMPLIANCE This instrument has been tested and found to be in compliance with all applicable requirements of the following safety and electromagnetic standards: • UL Std No. 61010A-1 Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1: General Requirements • UL Std No. 61010A-2-010 Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1: General Requirements • CAN/CSA C22.2 No. 1010.1-92 - Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1: General Requirements (includes Amendment 1) • CAN/CSA C22.2 No. 1010.1B-97 - Amendment 2 CAN/CSA C22.2 No. 1010.1-92 - Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1: General Requirements • CAN/CSA C22.2 No. 1010.2.010A-97 - Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 2-010: Particular Requirements for Laboratory Equipment for the Heating of Materials, Amendment No. 1 • IEC 61010-1 Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory Use, Part 1: General Requirements • IEC 61010-1 Safety Requirements for Electrical Equipment for Measurement, Control, and Laboratory use, Part 2: Particular Requirements for Laboratory Equipment for the Heating of Materials

ELECTROMAGNETIC COMPATIBILITY (EMC) • FCC Title 47 Part 15B as a Class A digital device • EN61326 Class A Electrical Equipment for measurement, control, and laboratory use - EMC Requirements

FCC WARNINGS AND NOTES • Warning. Changes or modifications to this unit, not expressly approved by the party responsible for compliance, could void the user’s authority to operate the equipment • Note. This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to part 15 of the FCC Rules. These limits are designed to provide

iv

S1000 Thermal Cycler Manual

reasonable protection against harmful interference when the equipment is operated in a commercial environment. This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the instruction manual, may cause harmful interference to radio communications. Operation of this equipment in a residential area is likely to cause harmful interference, in which case the user will be required to correct the interference, at his own expense • Note regarding FCC compliance. Although this design of instrument has been tested and found to comply with Part 15, Subpart B of the FCC Rules for a Class A digital device, please note that this compliance is voluntary, for the instrument qualifies as an “exempted device” under 47 CFR 15.103(c), in regard to the cited FCC regulations in effect at the time of manufacture • Note regarding Canadian EMC compliance: Le present appareil numerique n’emet pas de bruits radioelectrique depassant les limites applicables aux appareils numeriques de class A prescrites dans le reglement sur le brouillage radioelectrique edicte par le Ministere des Communications du Canada • Cables. Shielded cables must be used with this unit to ensure compliance with the Class A FCC limits

v

S1000 Thermal Cycler Manual

Table of Contents Bio-Rad Laboratories Resources... ii Warranty... ii Safety and Regulatory Compliance... ii

Chapter 1. Introduction to the S1000™ Thermal Cycler... 1 System Overview... 1 Reaction Modules... 2 Setting Up the S1000 Thermal Cycler... 3 Operating the Reaction Module Lid... 4 Main Menu... 5

Chapter 2. Creating and Editing Protocols... 7 Creating a New Protocol... 7 Editing an Existing Protocol... 10 Sample Volume and Lid Temperature... 12

Chapter 3. Running Protocols... 13 Preparing to Run a Protocol... 13 Monitoring the Protocol Run... 14 Canceling a Run... 15 Incubating Samples... 15

Chapter 4. Managing Protocol Files and Folders... 17 Managing Protocol Files and Folders... 17

Chapter 5. Advanced Tools and Functions... 21 TOOLS Options... 21 Controlling S1000 Thermal Cyclers with a C1000™ or C1000 TouchTM... 21

Chapter 6. Maintenance and Troubleshooting... 23 Cleaning and Maintaining the S1000 Thermal Cycler... 23 Maintaining Sufficient Airflow... 25 Replacing Fuses... 26

vii

S1000 Thermal Cycler Manual

viii

S1000 Thermal Cycler Manual

1

Introduction to the S1000™ Thermal Cycler Read this chapter for information on setting up the S1000 thermal cycler. • System overview (page 1) • Reaction modules (page 2) • Setting up the S1000 thermal cycler (page 3) • Operating the reaction module lid (page 4) • Main menu (page 5)

System Overview The S1000 thermal cycler base (Figure 1) includes: • Reaction module bay - holds the inserted reaction module • Reaction module locking bar - locks the inserted module in place • Control panel - provides access to all the functions needed to create and run PCR protocols • Air vents - allow the thermal cycler to heat and cool quickly Module locking bar Reaction module bay Control panel

Air vents

Figure 1. Front view of the S1000 thermal cycler. The back panel of the S1000 thermal cycler includes data ports (Figure 2). • USB B port - connects the S1000 thermal cycler to a C1000TM or C1000 TouchTM thermal cycler • USB A ports - inactive

1

S1000 Thermal Cycler Manual

• Test port - for service testing only USB B port

Test port

USB A ports Power switch Power input Fuses Cooling vents Figure 2. Back panel of the S1000 thermal cycler. The control panel on the S1000 thermal cycler provides access to all the functions needed to run the thermal cycler and includes the following components: • Liquid crystal display (LCD) - displays the main menu and other screens • Command, numeric, and navigation keys - use these keys to enter commands, numbers, or letters and to navigate various screens The main screen is displayed after booting is complete. Figure 3 shows the components of the control panel. Numeric and LCD command keys Main menu options Navigation keys Screen key

Figure 3. Components of the control panel on the S1000 thermal cycler.

Reaction Modules The S1000 thermal cycler is compatible with any 1000-series reaction module. The reaction modules come in four sizes: the 96-well fast, 96-deep well, dual 48/48-well, or 384-well blocks. Each reaction module includes a fully adjustable heated lid that is capable of running reliably with a broad range of reaction vessels. Each reaction module contains cooling fins for fast heating and cooling and a fully adjustable, heated lid. • Heated inner lid - adjusts the lid temperature to prevent condensation and evaporation • Sample/reaction block - holds reaction vessels, including tubes and microplates

2

S1000 Thermal Cycler Manual

The top of a reaction module lid includes a lid lever, lid force knob, and status LED. • Lid lever - opens and closes the lid • Lid force knob - sets lid force and seals the reaction • Status LED - turns on to indicate that the block is selected or running Figure 4 shows the components of a 96-well fast reaction module. Lid

Lid force knob Lid lever Status LED

Cooling fins

Figure 4. Components of a reaction module.

Setting Up the S1000 Thermal Cycler The S1000 thermal cycler package includes: • S1000 thermal cycler base • Power cord • Consumables selection guide • Instruction manual • Quick guide for system installation Reaction modules for use with the S1000 thermal cycler are shipped in separate packaging. Remove all packaging materials and store them for future use. If any item is missing or damaged, contact your local Bio-Rad office. Place the S1000 thermal cycler base on a flat, dry surface with sufficient cool airflow to run properly. The instrument can run in two modes: stand-alone or software-controlled. When running the system under software-controlled mode, make sure during setup there is sufficient space for a computer. To insert a 96-well fast, 96-deep well, dual 48/48-well, or 384-well reaction module into the reaction module bay of the thermal cycler base, follow these instructions: 1. With the locking bar in the down position and the lid lever of the reaction module pointing to the front, lift the reaction module into the reaction module bay. Leave about 1-2 cm of space in front of the module. 2. Pull the locking bar up to lock the reaction module in place. There is no space at the front of the module when it is locked into the S1000 thermal cycler base. 3. Plug the supplied power cord into the appropriate electrical outlet.

3

S1000 Thermal Cycler Manual

4. Turn on the thermal cycler using the power switch on the back panel of the thermal cycler base. NOTE: Before operating the thermal cycler, be sure to read the safety specifications (Safety and Regulatory Compliance on page ii) and operating requirements. 5. When the S1000 thermal cycler starts up, it goes through two screens: the black booting and the self-test screens. Once the self-test is run to verify proper functions, the main menu is displayed. Use the main menu to begin operating the thermal cycler. To remove the reaction module from the thermal cycler base, follow these instructions: 1. Turn off the thermal cycler. 2. Unlock and release the reaction module by pushing the locking bar down. 3. Carefully lift the reaction module out of the bay. WARNING! Cooling fins may be hot immediately after running a protocol or incubation. Before lifting the reaction module, make sure that the cooling fins are not hot. 4. After removing the reaction module from the S1000 thermal cycler, store it on a clean, flat surface where it cannot get bumped, scraped, or dropped.

Operating the Reaction Module Lid The inner lid of the reaction module applies heat and force to the reaction vessel lids (caps or tape) to produce consistent and successful reactions. Heating the inner lid prevents condensation, while applying force seals the reaction to prevent evaporation. WARNING! After a run, the heated inner lid can remain hot. Use caution when opening and closing the lid. To open the lid, use the following steps: 1. Turn the lid force knob counterclockwise to release the inner lid. 2. To open the lid, lift the lid lever up. 3. Lift the lid lever completely until the reaction module stays open without assistance. To close the lid, use the following steps: 1. Push the lid lever down, making sure that the front of the lid is secured beneath the housing. 2. Adjust the lid force by turning the lid force knob. • Turn the knob clockwise to increase the lid force • Turn the knob counterclockwise to decrease the lid force 3. Increase the lid force until the heated lid touches the reaction vessel. Continue to increase the lid force based on the following recommendations: • If using tubes, increase the lid force by a quarter turn after the lid touches the tubes • If using plates, increase the lid force by a half turn after the lid touches the plate NOTE: The position marks on the lid indicate quarter turns. WARNING! If the lid is tightened past the recommended force, tightening the lid further will result in the knob slipping and causing a clicking sound. This is designed as a warning that the lid force is past the recommended setting and may

4

S1000 Thermal Cycler Manual

result in damage to the reaction vessels. If this happens, decrease the lid force by turning the lid force knob counterclockwise 1 full turn. Re-apply the correct lid force by following the instructions above.

Main Menu The main menu (Figure 5) provides access to all thermal cycler operations and displays the status of the reaction module and the name of the thermal cycler. Name of thermal cycler Options

Status message Figure 5. The main window of the S1000 thermal cycler. Select the options in the main menu to start these instrument functions: • RUN - to run an existing protocol file • NEW - to create a new protocol file • EDIT - to modify stored protocol files • FILES - to copy, move, rename, delete, or secure protocol files and/or folders • VIEW - to review an existing protocol file • TOOLS - to change thermal cycler settings or to view the last protocol that was run

5

S1000 Thermal Cycler Manual

6

S1000 Thermal Cycler Manual

2

Creating and Editing Protocols Read this chapter for information on creating and editing protocols. • Creating a new protocol (page 7) • Editing an existing protocol (page 10) • Sample volume and lid temperature (page 12)

Creating a New Protocol NOTE: The internal memory of the S1000™ thermal cycler can hold up to 400 two-step protocols. To create a protocol: 1. Select NEW from the main menu. Press ENTER to confirm the selection. 2. Use the numeric keys to enter the name of the new protocol file. Enter a letter by pressing the up or down arrow key and a number by pressing the numbered key. For example, to select the letter C, press the up key 3 times. To select the letter S, press the down key eight times. Press ENTER to continue to the next space. Press ENTER to continue to the next screen. NOTE: A protocol file name can contain 1–8 characters and must be unique to the folder. To delete or change a letter, press CANCEL and select a new letter. To delete the entire name, press CANCEL multiple times. In Figure 6, the characters STD3 are entered, and the cursor is highlighting the next space.

Figure 6. STD3 is entered as the protocol name.

7

S1000 Thermal Cycler Manual

3. Using the arrow keys, select TEMP to enter a temperature step or GRAD to enter a gradient temperature step in the protocol file. Press ENTER to continue to the next screen. NOTE: The first step in a protocol must be either a TEMP or GRAD step. 4. Enter the target temperature between 0 (zero) and 100.0°C for the temperature step. Press ENTER to continue to the next screen. 5. Enter the hold time (TIME) in minutes and seconds using the numeric keys. The hold time (TIME) ranges between 0:01 (one second) and 18:00:00 (18 hours). Entering 0 (zero) adds an infinite hold and holds this step forever. Press ENTER to continue to the next screen. 6. Select YES, NO, or OPTION by pressing the right and left arrow keys. • YES - to confirm the current parameters for this protocol step • NO - to change a parameter in this protocol step • OPTION - to add more parameters to this protocol step. For more information about entering options, see Adding an Option to a Temperature Step (page 10) Press ENTER to continue to the next screen. 7. Enter the lower temperature in the gradient. The lower temperature is at the front (row H) of the block. 8. Enter the upper temperature in the gradient. The upper temperature is at the back (row A) of the block. NOTE: The range of temperatures is limited by the widest available range for gradient, which is 24°C. The highest value that can be entered for the upper temperature is 100°C. 9. Enter a hold time between 0:01 (one second) and 18:00:00 (18 hours). 10.Select YES, NO, or OPTION by pressing the right and left arrow keys. • YES - to confirm the current parameters for this protocol step • NO - to change a parameter in this protocol step • OPTION - to preview the temperature gradient. If Option is selected, select VIEW on the next screen to view the gradient or EXT to add a hold time extension. Press ENTER to continue to the next screen. 11.Repeat the instructions in steps 5–8 to continue entering additional temperature steps. NOTE: A protocol can contain up to 99 protocol steps. The first step must be a temperature (TEMP) step, while the last step must be an END step. 12.(Optional) To enter a GOTO step immediately after the set of steps to be repeated in a cycle, use the arrow keys and select GOTO. Press ENTER to continue to the next screen.

8

S1000 Thermal Cycler Manual

In Figure 7, step 5 is a GOTO step.

Figure 7. A GOTO step is selected. NOTE: The GOTO step cannot be the first or the last step in the protocol. 13.Enter the step number for the first step in the GOTO repeats using the numeric keys. Press ENTER to continue to the next screen. 14.Enter the number (between 1 and 9999) for the additional repeats (ADDTNL REPEATS) in the GOTO step. Press ENTER to continue to the next screen. NOTE: The GOTO step adds additional cycles to the PCR protocol. The first cycle is not included in the GOTO step. For example, to run a PCR protocol with 31 cycles, enter 30 repeats in the GOTO step. In Figure 8, the number of repeats is 30, and the total number of cycles is 31.

Steps that repeat in the GOTO step

Number of additional repeats in a GOTO step

Figure 8. Enter the number of repeats in a GOTO step. 15.Select YES to accept the GOTO step parameters, or select NO to return to the beginning of this step and change the GOTO step parameters. Press ENTER to continue to the next screen. 16.Enter the remaining steps by choosing the step type and adding parameters. Press ENTER to continue to the next screen. 17.Select END using the arrow keys to instruct the thermal cycler to finish the protocol file. Press ENTER to continue to the next screen. NOTE: The END step must be the last step of a protocol; a protocol can contain only one END step. 18.Select YES to accept the protocol step parameters or NO to return to the beginning and select a different protocol step. 19.Use the arrow keys to select the folder where you want the new protocol file to be saved and press ENTER to save the protocol file. NOTE: The file tree folder screen does not appear if there are no user-created folders (i.e. the MAIN folder is the only folder on the list). Saving the protocol in the MAIN folder is not recommended. If the protocol is saved in the MAIN folder, we recommend moving it to a user-created folder. (See Moving a Protocol File on page 18 for more information.)

9

S1000 Thermal Cycler Manual

Adding an Option to a Temperature Step To add an option: 1. Select OPTION using the arrow keys. Press ENTER to continue to the next screen. 2. Select the option you want to add to the protocol step using the arrow keys. Press ENTER to continue to the next screen. 3. Enter the option value using the numeric keys. 4. To confirm the parameters of the protocol step, select YES and then press ENTER. To change the parameters, select NO and then press ENTER.

Editing an Existing Protocol NOTE: A protocol that is already running cannot be edited. Changes made in a protocol that is running apply to the next time the protocol runs. To stop editing a protocol, press CANCEL several times.

Editing the Lid Temperature and Sample Volume To edit an existing protocol: 1. Select EDIT from the main menu. Press ENTER to confirm the selection. 2. Using the arrow keys, select the folder that contains the protocol file to be edited. Press ENTER to continue to the next screen. 3. Enter the new lid temperature (optional) or use the default lid temperature. Press ENTER to accept the lid temperature and continue to the next screen. NOTE: The lid temperature can range from 0 to 110°C. When the block is running an infinite hold at a temperature below the Turn off below parameter, the lid heater maintains 31.0°C. To change the default Turn off below parameter, select TOOLS > DEFAULTS. 4. Enter a new sample volume (optional) or use the default volume. Press ENTER to continue to the next screen.

Inserting a Protocol Step 1. Select EDIT from the main menu. Press ENTER to confirm the selection. 2. Using the arrow keys, select the folder that contains the protocol file to be edited. Press ENTER to continue to the next screen. 3. Select a protocol step to edit using the arrow keys. Press ENTER to continue editing the step. 4. Select INS to insert a step above the selected protocol step. 5. Select TEMP, GRAD, or GOTO as the type of protocol step to be inserted. Press ENTER to continue to the next screen. 6. Enter the step parameters. Press ENTER to confirm each parameter. 7. Enter the parameters of the new step. Press ENTER to confirm each parameter.

10

S1000 Thermal Cycler Manual

Deleting a Protocol Step 1. Select EDIT from the main menu. Press ENTER to confirm the selection. 2. Using the arrow keys, select the folder that contains the protocol file to be edited. Press ENTER to continue to the next screen. 3. Select a protocol step to delete using the arrow keys. Press ENTER to continue editing the step. 4. Select DEL to delete the selected protocol step. Press ENTER to continue to the next screen. Notice that the deleted step parameters are replaced with the parameters of the next step. 5. Confirm the deletion. When prompted with Save changes?, select YES and press ENTER to delete the step. Alternatively, select NO and press ENTER to return to the beginning of this step.

Editing a Protocol Step To change the parameters in existing protocol steps, use the following instructions: 1. Select EDIT from the main menu. Press ENTER to confirm the selection. 2. Using the arrow keys, select the folder that contains the protocol file to be edited. Press ENTER to continue to the next screen. 3. Select a protocol step to delete using the arrow keys. Press ENTER to continue editing the step. 4. Select EDIT to delete the selected protocol step. Press ENTER to continue to the next screen. TIP: When you first select EDIT, you can edit the parameters in the selected protocol step. To edit the parameters in a different protocol step, press the arrow keys to go to that step. 5. Change the first parameter in the step by pressing the keys on the control panel to enter the temperature. Press ENTER to continue to the next parameter. 6. Change the second parameter in the step by pressing the numeric keys to type a new parameter. Press ENTER to continue to continue to the next step. 7. Continue editing the existing parameters in each step (optional). Press the up and down arrow keys to choose an earlier or later step in the protocol. In Figure 9, the temperature parameter is changed to 70.0°C in step 4.

Enter new parameter in next step Figure 9. The temperature parameter is changed to 70.0°C.

11

S1000 Thermal Cycler Manual

8. Press ENTER to finish changing parameters in the step and to continue editing the protocol.

Sample Volume and Lid Temperature The sample volume and lid temperature influence the outcome of a PCR protocol. • Sample volume - determines the temperature control mode, which influences the amount of time the samples are held at the target temperature • Lid temperature settings - determine the temperature of the heated lid and when it cuts off. If the temperature is too high, the sample temperature might rise above the target temperature

Temperature Control Modes The S1000 thermal cycler uses one of two temperature control modes to determine when the sample reaches the target temperature: • Calculated mode - The thermal cycler calculates the sample temperature based on the sample volume when a sample volume between 1 and 50 μl is entered for 96well or dual 48/48-well reaction modules, a volume between 1 and 30 μl is entered for the 384-well reaction module, or a volume between 1 and 125 μl is entered for the 96-deep well reaction module. The calculated mode is recommended, because it most accurately represents the actual sample temperature • Block mode - When a sample volume of zero (0) μl is entered, the thermal cycler assumes that the sample temperature is the same as the measured block temperature

12