TC-5000 OPERATOR'S MANUAL Issue 1 5/09
CONTENTS PAGE INTRODUCTION 3 SAFETY AND INSTALLATION INFORMATION Operator Safety Warning Unpacking Installation Replacement Cable Working Conditions Guarantee 4 4 4 4 4 5 5 6 SÉCURITÉ ET CONSIGNES D'INSTALLATION Sécurité de l'opérateur Avertissement Déballage Installation Câble de rechange Conditions de travail Garantie 7 7 7 7 7 8 8 9 SICHERHEITS - UND INSTALLATIONSINFORMATIONEN Sicherheit des Bedienpersonals Achtung Auspacken Inbetriebnahme Ersatzkabel Einsatzbedingungen Garantie 10 10 10 10 10 11
INSTRUMENT CONNECTIONS 19 CONTACT INFORMATION 20 TC-5000 SPECIFICATION 21 TC-5000 OPERATION AND MAINTENANCE Gradient Adjustable Heated Lid Adjustable Screen Consumables PC Connections Cleaning the TC-5000 Removing the Block 23 23 24 25 25 26 27 28 PROGRAMMING THE TC-5000 Start up Menu The System User Menu Writing and Editing Programs Option 1: Copying an Existing Template Option 2: Editing and Saving an Existing Template Option 3: Creating a New Program Inserting a Segment Inserting Cycle Numbers In
INTRODUCTION The TC-5000 is a gradient thermal cycler, designed to accurately control the temperatures of samples between 4°C and 99°C. It has many scientific applications including DNA amplification, PCR 1, reverse transcription and sequencing. The TC-5000 is programmed via a touch screen LCD graphics display.
SAFETY AND INSTALLATION OPERATOR SAFETY Please read this manual carefully before using the thermal cycler. If the equipment is not used in the manner described in this manual the protection provided by the equipment might be impaired. All Bibby Scientific Ltd instruments are designed to conform to international safety requirements and are fitted with an over-temperature cut-out.
rating plate (located on the back of the unit). Ensure that the voltage selector switch (located above the On/Off switch) is set to the appropriate voltage for the local supply. Please note that the unit must be earthed to ensure proper electrical safety. Connect the mains cable to a suitable plug according to the table below. Connections Live Neutral Earth 220V-240V, 50/60Hz Supply brown blue green/yellow 110V-120V, 50/60Hz Supply black white green 3.
GUARANTEE The instrument is guaranteed against any defect in material or workmanship for the period specified on the enclosed guarantee card. This period is effective from the date of purchase; within this period all defective parts will be replaced free of charge provided that the defect is not the result of an accident, misuse or negligence. The screen of the thermal cycler can be damaged by sharp objects such as pens, pencils and fingernails.
SÉCURITÉ ET CONSIGNES D'INSTALLATION SÉCURITÉ DE L’OPÉRATEUR Lire attentivement le présent manuel avant d'utiliser le cycleur thermique. Si ce matériel n'est pas utilisé de la manière décrite dans le présent manuel, la protection fournie risque d'être compromise. Tous les appareils Bibby Scientific Ltd sont conçus pour être conformes aux exigences internationales de sécurité et sont dotés d'une coupure en cas de surchauffe.
2. Avant de raccorder l'appareil à l'alimentation électrique sur secteur, vérifier la tension par rapport à la plaque signalétique (située au dos de l'appareil). Vérifier que le sélecteur de tension (situé audessus de l'interrupteur Marche/Arrêt) est réglé sur la tension correspondant au réseau local. Noter que l'appareil doit être relié à la terre pour assurer une bonne sécurité électrique. Brancher le câble secteur sur une fiche adéquate d'après le tableau suivant.
L'appareil a été testé en matière de perturbations radioélectriques et est certifié selon la norme EN61326. GARANTIE L'appareil est garanti contre tout défaut de matériaux ou vice de fabrication pendant la période précisée sur la carte de garantie jointe. Cette période s'applique à compter de la date d'achat; pendant cette période, toutes les pièces défectueuses seront remplacées gratuitement à condition que le défaut ne soit pas le résultat d'un accident, d'une mauvaise utilisation ou d'une négligence.
SICHERHEITS - UND INSTALLATIONSINFORMATIONEN SICHERHEIT DES BEDIENPERSONALS Lesen Sie diese Anleitung vor Verwendung des Thermocyclers bitte sorgfältig durch. Wenn das Gerät nicht entsprechend der Bedienungsanleitung eingesetzt wird, können die Schutzfunktionen des Gerätes beeinträchtigt werden. Alle Geräte von Bibby Scientific Ltd entsprechen den internationalen Sicherheitsanforderungen und sind mit einer Übertemperatur-Schutzvorrichtung ausgestattet.
1. Alle Geräte von Bibby Scientific Ltd werden mit einem Netzkabel geliefert, das entweder eingesteckt wird oder fest mit dem Gerät verbunden ist. 2. Vor dem Anschluss bitte kontrollieren, ob die Stromversorgung den Angaben auf dem Typenschild (auf der Geräterückseite) entspricht. Darauf achten, dass der Spannungswähler (über dem Ein/AusSchalter) auf die entsprechende örtliche Spannung eingestellt ist. Um die elektrische Sicherheit zu gewährleisten, muss dieses Gerät geerdet werden.
Hinweis: Die Gerätespezifikationen beziehen sich auf eine Umgebungstemperatur von 20°C und können sich außerhalb des Bereichs 10°C bis 30°C verschlechtern. Das Gerät wurde auf HF-Störeinflüsse geprüft und entspricht den EMV-Bedingungen nach EN61326. GARANTIE Bibby Scientific Ltd gewährleistet, dass dieses Gerät für den auf der Garantiekarte angegebenen Zeitraum keine Herstellungs- und Materialmängel aufweist. Dieser Zeitraum tritt ab dem Verkaufsdatum in Kraft.
INFORMACIÓN DE SEGURIDAD E INSTALACIÓN SEGURIDAD DEL OPERARIO Lea atentamente este manual antes de utilizar el ciclador térmico. Si el equipo no se utiliza de la forma descrita en este manual, se reducirá la protección ofrecida por el equipo. Todos los instrumentos Bibby Scientific Ltd están diseñados para cumplir los requisitos internacionales de seguridad, e incluyen un dispositivo de corte de sobretemperatura.
2. Antes de conectar el instrumento al suministro eléctrico, compruebe que el voltaje coincida con el indicado en la placa de régimen (situada en la parte trasera de la unidad). Asegúrese de que el interruptor de selección de voltaje (situado encima del interruptor de encendido/apagado) está ajustado al voltaje correcto del suministro eléctrico. El instrumento debe disponer de una toma de tierra para garantizar la seguridad eléctrica adecuada.
especificaciones pueden empeorar si se utiliza el instrumento fuera del intervalo de temperatura comprendido entre 10°C y 30°C. Se han realizado pruebas para comprobar la interferencia de radiofrecuencia del instrumento, el cual cumple la normativa EN61326. GARANTÍA El instrumento está garantizado contra cualquier defecto en el material o la fabricación durante el período especificado en la tarjeta de garantía que se adjunta. Este período entra en vigor a partir de la fecha de compra.
INFORMAZIONI SULLA SICUREZZA E L'INSTALLAZIONE SICUREZZA DELL'OPERATORE Leggere attentamente il presente manuale prima di usare il ciclatore termico. Se non si usa l'apparecchiatura nel modo descritto nel presente manuale, la protezione fornita dall'unità potrebbe risultarne diminuita. Tutti gli strumenti Bibby Scientific Ltd sono progettati per rispettare i requisiti di sicurezza internazionali e sono montati con un sezionatore di sovratemperatura.
2. Prima di collegare lo strumento all'alimentazione elettrica di rete, controllare la tensione confrontandola con la targhetta riportante i valori nominali (si trova sul retro dell'unità). Assicurarsi che il selettore di tensione (posto sopra l'interruttore On/Off) sia impostato sulla tensione corretta per l'alimentazione locale. Notare che al fine di garantire la corretta sicurezza elettrica, occorre che l'unità sia messa a terra.
Nota: le specifiche di controllo sono indicate ad una temperatura ambiente di 20°C. Le specifiche potrebbero peggiorare fuori da una temperatura ambiente compresa tra 10°C e 30°C. Lo strumento è stato collaudato per interferenze da radiofrequenze ed è certificato secondo la norma EN61326. GARANZIA Lo strumento è garantito da qualsiasi difetto nei materiali o nella lavorazione per il periodo specificato nella scheda di garanzia allegata.
INSTRUMENT CONNECTIONS Serial Port RS232/RS485 Selector: Select RS485 for multiple units or RS232 for a single unit Parallel Port P RS 485 A R A RS485 Connections L RS 485 L E RS 485 S E R RS 232 I A L L D AISY TERMINATION CHAIN BOX Connection Switch: Select “Daisy Chain” for all units except the last in the chain and “Termination Box” for the last unit Voltage Selector: Select 230V or 115V and ensure correct fuses are fitted On/Off Switch Power Lead Connection: Ensure that appropriate
CONTACT INFORMATION For technical, sales or servicing information, contact your local Bibby Scientific Ltd dealer or: North and South America Techne Inc. 3 Terri Lane, Suite 10, Burlington, NJ, 08016, USA Toll free: 800-225-9243 Tel: +1 609-589-3560 Fax: +1 609-589-2571 e-mail: labproducts@techneusa.com www.techneusa.com UK and Rest of World Bibby Scientific Ltd Beacon Road, Stone, Staffordshire, ST15 0SA, UK Tel: +44 (0)1785 812121 Fax: +44 (0)1785 813748 e-mail: techne@bibby-scientific.com www.
TC-5000 SPECIFICATION Technical Specification at an ambient temperature of 20°C Temperature Gradient Range Maximum Gradient Minimum Gradient Number of temperature probes Block temperature range Typical heating rate Typical cooling rate Block uniformity (without gradient) Column uniformity with a 10°C gradient Control accuracy Gradient accuracy Temperature set point adjustment 20°C to 70°C 30°C 1°C 4 4°C to 99°C 3.0°C/sec 1.3°C/sec <+ _ 0.3°C <+ _ 0.3°C <+ _ 0.3°C + < _ 0.3°C + _ 0.
Miscellaneous Height x Width x Length Weight Power consumption Parallel port Serial port RS232/RS485 adaptable Connection to PC 260mm x 220mm x 420mm 12.
TC-5000 OPERATION AND MAINTENANCE The TC-5000 has many features which increase the flexibility of the instrument and improve the ease of use. The following features are described in more detail in this chapter: • • • • Gradient Adjustable heated lid Adjustable screen PC connections This chapter also includes useful information relating to the type of consumables which can be used in the thermal cycler.
It is important to remember that the greater the gradient, the poorer the uniformity across the block, therefore fine-tuning may be required to establish the optimum temperature. While the gradient is mainly linear there are small variations at the extremes of the gradient due to a loss of heat from the hottest part of the block and a gain of heat from the adjacent column on the coldest.
TC -5 0 00 A heated lid mat (Part No. 6104978) is supplied with the TC-5000; this is for use with 96-well PCR plates sealed with an adhesive seal. The rubber mat should be placed in top of the sealing film to ensure good thermal contact with the heated lid. Do not over-tighten the lid as this will cause the silicone rubber membrane to deform and will reduce the effectiveness of the heated lid.
PC CONNECTIONS It is possible to connect either a single thermal cycler or a series of instruments to a computer. The options are: 1. To connect a single thermal cycler to a computer part number FGEN232 is required which contains a connecting cable, software disk and instructions. 2. To connect several units via a daisy chain to a single computer part number FTGEN485 is required which contains an RS485 set.
CLEANING THE TC-5000 Prior to cleaning the thermal cycler it is important to disconnect the unit from the power supply and if it has been in use allow both the block and heated lid to cool to below 50°C prior to attempting to clean it. The peltier block, in particular the sample wells should be cleaned regularly to ensure that the maximum heat transfer from the block to the sample occurs. The block should be cleaned if a spillage occurs before the instrument is used again.
REMOVING THE BLOCK The thermal cycling block can be removed for detailed cleaning or replacement. First ensure that the unit is disconnected from the power supply and has been allowed to cool if the unit was previously in use. The two quick release handles located at the front and back of the block are used to unlock the block. The padlock symbols indicate the direction in which to slide the handles: open to unlock and closed to lock. When the block is unlocked it will simply lift out of the unit.
PROGRAMMING THE TC-5000 START UP MENU When the TC-5000 is first switched on the screen shown below is displayed and will remain until the unit has finished performing a series of self tests. This screen displays information such as the serial number of the instrument, the type of block being used and the version of software running the thermal cycler. Techne TC-5000 Serial number Software version Hardware version Block ID Last system test Comms address Copyright protected 2003 xxxxxx/xx v.vv h.
THE SYSTEM USER MENU In this section of the software the user is able to write new programs and edit existing ones. Additionally the program defaults for parameters such as the heated lid and final extension steps can be altered. The gradient calculator and data log files can also be accessed through their sub-menus. To access the programming software press the "System User" button on the start-up screen. The System User menu will then appear.
WRITING AND EDITING PROGRAMS There are three options available for creating programs on the TC-5000: 1. Copy and edit one of the pre-written templates. 2. Edit a pre-written template and save it using a different name. 3. Create a new program with user-specific parameters. There are 5 pre-written templates. These templates cannot be altered; however, they can be copied, given a new name and then edited to save time when writing a new program.
OPTION 1: COPYING AN EXISTING TEMPLATE Techne TC-5000 To access the program templates select "Programs" in the System Users menu. Programs Program Defaults Gradient Calculator Exit The cursor arrows can be used to scroll up and down the stored programs. An individual program is opened by pressing on the individual program box (shown opposite). Select Program 14526 bytes of memory free b 2 STEP PCR TEMPLATE b 3 STEP PCR TEMPLATE Exit The programs are pre-fixed by b or i.
Select the copied program. Select Program 14526 bytes of memory free b ICE BUCKET b LITIGATION 15 DEGREES b RT PCR TEMPLATE i B ACTIN i GAPDH ▼ ▼ New Program Exit The program cannot be changed at this point so press "Edit" to make any changes required. B ACTIN Last Modified 15/05/2006 Heated lid 11:29:17 105.0°C Heated lid before program On Pause before program Off Initial denaturation 05m00s The program can be copied, deleted, printed or run by selecting the appropriate option. 94.
OPTION 2: EDITING AND SAVING AN EXISTING TEMPLATE Select Program 14526 bytes of memory free Select the template from the program menu. b 2 STEP PCR TEMPLATE b 3 STEP PCR TEMPLATE b ICE BUCKET b LITIGATION 15 DEGREES b RT PCR TEMPLATE ▼ ▼ New Exit Select "Edit" Program 3 STEP PCR TEMPLATE Last Modified 01/11/1998 00:00:00 Heated lid 105.0°C Heated lid before program On Pause before program Off Initial denaturation 05m00s 94.
OPTION 3: CREATING A NEW PROGRAM Select Program 14526 bytes of memory free The final option is to create a completely new program by selecting "New" in the Select Program menu. This section also contains details of the individual parameters and functions. b 2 STEP PCR TEMPLATE b 3 STEP PCR TEMPLATE b ICE BUCKET b LITIGATION 15 DEGREES b RT PCR TEMPLATE To create a new program, select "New" in the Select Program menu. ▼ ▼ New Editing Type in a program name; for details see the previous page.
The next step enables one of the following to be added to the program: Edit program function Next step Segment 7 8 9 ➔ Number Cycles 4 5 6 Continue Inc Dec 1 2 3 Cancel Pause 0 • BS OK Segment: Enables the temperature, time and heating rate to be inserted into the segment. Additionally, a gradient across the block can be added. ➔ Number Cycles: Up to 99 cycles can be added Inc Dec: Allows increasing and decreasing temperatures and times to be set. See page 37 for further details.
The time can be set in hours (h), minutes (m) or seconds (s). Type in the time required followed by the unit. Use the ➔ to move across the screen to the ramp rate. Edit program function Segment 94.0°C 0°C 00m30s MAX 0°C/s Segment h 7 8 9 ➔ Number Cycles m 4 5 6 Continue Inc Dec s 1 2 3 Cancel 0 • BS OK Pause ➔ The ramp rate is automatically set to the maximum 3°C per second. A number less than 2°C/s can be inserted. Selecting "Max" will reset the ramp to the maximum.
Temperature Cycle 1 Cycle 2 Cycle 3 Cycle 4 Decremental temperature for Touchdown DNA amplification -i -i -i Time The Hold Temperature of the incremented/decremented step is the defined Hold Temperature plus the summation of the increments/decrements. The first cycle is never altered, only subsequent cycles. If decrement is selected, the Hold Temperature is prevented from falling below 4°C. If increment is selected the Hold Temperature is prevented from rising above 99°C.
INSERTING A PAUSE Selecting "Pause" adds a stop to the program, which when running requires user intervention before the program can continue. Edit program function PAUSE Segment 7 8 9 ➔ Number Cycles 4 5 6 Continue Inc Dec 1 2 3 Cancel Pause 0 • BS OK ➔ SAVING CHANGES TO A PROGRAM Editing To change a saved program, select the program and when opened select "Edit" to change the parameters. B ACTIN Heated lid 105.
Select the program required, using the cursor keys to view all the available programs. Select Program 14526 bytes of memory free b 2 STEP PCR TEMPLATE b 3 STEP PCR TEMPLATE b ICE BUCKET b LITIGATION 15 DEGREES b RT PCR TEMPLATE ▼ ▼ New Exit Press "Run". Program 3 STEP PCR TEMPLATE Last Modified 01/11/1998 Heated lid 00:00:00 105.0°C Heated lid before program On Pause before program Initial denaturation Off 05m00s 94.
As the program continues the temperatures are displayed on the screen, as shown opposite. B ACTIN Program Cycle 3 of 3. Step 2 of 3 Holding at 55.0°C 10°C Sample Lw= 50.0°C, Sample Up= 60°C Time to complete 01m26s 100 50 0 00h01m 00h02m Stop 00h03m 00h04m 00h05m Pause Program Program complete Sample = 72°C Time to complete 00m00s This screen is displayed when the run is complete. If there is a final hold in the program press "Stop" to end the program.
100 This screen is displayed if there is a power loss during the running of the program and the auto-restart function is switched on. The display shows the times at which the power failed and resumed. 50 The "Show Info" button takes the screen back to the program information and display.
Pause before program: This allows the insertion of a pause in the experiment before the experiment starts. After the pause there must be user intervention before the program can move on to the next step of the program. Initial denaturation: Many experimental protocols require a high temperature incubation to remove any secondary structure of the nucleic acid template.
GRADIENT CALCULATOR The gradient is set within the program (see page 36 for further details), however, it is not possible to view the actual temperatures of each column in the program. The gradient calculator calculates the temperatures for each of the 12 columns (24 for the 384 well block) and allows the user to determine the optimal amplification conditions for the assay. The Gradient Calculator is found in the System User menu.
SYSTEM ADMINISTRATION This section of the software contains settings for the instrument, rather than the actual programs. From various sub-menus it is possible to change passwords for programs and the instrument, test the instrument, delete current programs stored on the memory card and completely reset the instrument. To access the System Admin simply press the "System Admin" button on the start-up screen.
The following functions are located within the System Administration menu: Program Passwords: Enables an administrator to change the password for any program. Sys Admin Password: Allows the administrator to change the administrator password. System Test: Initiates a simple diagnostic test. Controller Reset: Allows the user to restore the factory settings. System Configuration: This sub-menu allows the user to change the format of dates, times and temperature units.
SYSTEM TEST This is a diagnostic test to ensure that the Real Time Clock settings are correct. Techne TC-5000 RTC OK Exit CONTROLLER RESET If "Confirm Reset" is pressed the instrument default settings will be restored. Techne TC-5000 WARNING! All files and programs which have been stored on the internal memory will be lost. *** Controller Reset *** WARNING!! This will reset the unit to system defaults. All stored files will be lost.
SYSTEM CONFIGURATION This contains all the settings for programming the instrument such as units of temperature, date format, language, and date and time settings. Other important features are enabling the auto restart in case of a power failure and setting the comms address of the instrument if multiple units are linked to a computer. This part of the software also contains information relating to the cycle count for the block and the heated lid.
Date: The date can be corrected by the user by using the numerical interface shown above. Time: The time can be corrected by the user by using the numerical interface shown in above. Auto restart: This allows a program to be re-started in the event of a power failure when the instrument is in the middle of running a program. The options are Enabled or Disabled. Baud Rate: This is the speed of communication between the thermal cycler and a computer.
FREQUENTLY ASKED QUESTIONS Q1 What is "Hot start"? A1 The Hot Start step is used to pause the machine at a specific temperature, typically around 70°C, after the initial template denaturation. The reason is to allow the manual addition of unmodified Taq DNA polymerase which may loose activity if added during the initial 5min denaturation. Heat-activated Taq or Hot Start enzymes do not require this step. Q2 What is the incremental function? (See page 37).
ADDITIONAL INFORMATION Brief fault finding notes and a list of replacement parts are given in this section. Note: that this equipment should only be dismantled by properly trained personnel. Removing the outer cover exposes potentially lethal mains voltages. There are no user serviceable parts within this equipment. FAULT FINDING Should you have any problems with your TC-5000 which cannot be easily remedied, you should contact your supplier and return the unit if necessary.
Product Number Description Quantity FHSEALSD Heat sealer 1 FHSPA96 Heat sealer plate adapter for 96-well plates 1 FHSPA384 Heat sealer plate adapter for 384-well plates 1 FTC51BHD TC-5000 block for 96 x 0.2ml microtubes or 96-well plate 1 TC-5000 block for 96 x 0.2ml microtubes or 96-well fully skirted plate 1 FTC51B5D TC-5000 block 60 x 0.
Bibby Scientific France SAS Bibby Scientific Italia Srl ZI du Rocher Vert - BP 79, 77793 Nemours Cedex, France Tel: +33 1 64 45 13 13 Fax: +33 1 64 45 13 00 e-mail: bsf@bibby-scientific.fr www.bibby-scientific.com Via Alcide de Gasperi 56, 20077 Riozzo di Cerro al Lambro, Milano, Italia Tel: +39 (0)2 98230679 Fax: +39 (0)2 98230211 e-mail: marketing@bibby-scientific.it www.bibby-scientific.it Bibby Scientific Ltd Techne Inc.