PCR Thermal Cycler Calibration Using MTAS

Why Thermal Cycler Calibration?
Thermal cycler calibration is essential to improve PCR yields, control reagent usage and conform to regulatory requirements.

Critical decisions are often made on the outcome of PCR (Polymerase Chain Reaction) and qPCR reactions, particularly when applied in clinical, forensic or diagnostic applications. Major variables like thermal cyclers, reagents and disposables can have a significant contribution to and impact on the final results. It is essential to monitor performance of thermal cyclers as any variation can substantially affect the PCR process.

  • MultiProbe dynamic measurements
  • UKAS accredited
  • Brand independent
  • Wireless systems to validate qPCR & RT thermal cyclers including Light Cyclers
  • On-site calibration for minimal disturbance and downtime
  • Everything you need to know: uniformity, accuracy, ramp speeds, overshoots, undershoots, hold time
     

Thermal Cycler Calibration Service
METTLER TOLEDO offers the most sophisticated and accurate temperature calibration service for your PCR thermal cycler with UKAS accreditation. CYCLERtest provides assurance, with detailed information on how your PCR machine is performing throughout a reaction. Reporting on the real time performance of your PCR machines helps you better understand your results, adapt protocols for more efficient use of valuable reagents and conform to regulatory requirements.

  • For a quote, use the Request Service button at the end of this page.
     

Forensic and Diagnostic Applications
For forensic and diagnostic applications complete confidence in the procedure and data obtained is essential and can only be guaranteed with regular checking of the instrument used. UKAS accreditation of this system ensures you are complying to the most stringent laboratory practices of ISO:17025, whether you work to these guidelines or not.


Reliable PCR Results
The use of biological positive, negative and threshold controls is insufficient to guarantee reliable PCR results but the unique method of CYCLERtest performs both static and dynamic testing on every commercially available PCR thermal cycler. CYCLERtest uses the MTAS® (Mobile Temperature Acquisition System) that has been calibrated by KEMA (recognised internationally and by UKAS). It is verified by a “Council for Accreditation”.
 

Encrypted Test Results
Test results are analysed and reported through an automated process, prohibiting any alterations of the acquired sample data. The Data and layouts are encrypted and cannot be modified. A “Certificate of Thermal Calibration” accompanies every test report. This certificate represents the results of the measured and calculated main parameters of the tested instrument.
 

Trained and Certified Technicians
METTLER TOLEDO's policy is that only trained and certified technicians perform these calibrations and certifications. A technician will come to your laboratory for maximum efficiency and convenience and test your thermal cyclers, providing a full report on their functionality. A full understanding of your thermal cycler’s performance will improve optimisation of PCR reactions.
 

Independent Calibration of All Makes and Models
METTLER TOLEDO offers UKAS accredited calidation on all makes and models of thermal cycler including Real-Time PCR Machines. You can find the complete listing on CYCLERtest.
 

DRIFTCON End User Thermal Cycler Calibration Systems
Besides its calibration services, METTLER TOLEDO also provides DRIFTCON an end user calibration system which allows laboratories to perform in house calibrations.
 

FAQs

CYCLERtest FAQs

1. Can METTLER TOLEDO test all brands of thermal cyclers?
Yes. CYCLERtest is a completely independent calibration system and as long as your thermal cycler has a regular 0.2ml, 0.5ml or 384 well block METTLER TOLEDO can carry out the calibration. We can also test gradient cyclers with our special gradient protocol, and multiple and/or exchangeable blocks.

2. Can CYCLERtest be used to calibrate my Real Time PCR machine?
Yes, there is a new wireless version of CYCLERtest which has been specifically designed to measure the closed (q)PCR/RT-PCR system. It works by sending the results to the hardware box via radio waves. This system is compatiable with the ABI 7500, ABI 7300, ABI 7900HT with standard 96 well format and Stratagene MX3000 and MX3005P systems. Roche light cyclers can also be calibrated.

3. Can temperature deviations really influence my PCR results?
Yes, when the denaturation temperature is too high, the DNA polymerase will soon loose activity during the PCR process, resulting in low or no yields. Annealing temperatures are very specifically determind. Changes will result in specific bands or no annealing at all./p>

4. How can I change the standard 8 step protocol of my Driftcon system?
By registering on the Driftcon web site www.driftcon.com you can obtain up to eight different protocols free of charge.

5. How does the heated lid affect the CYCLERtest calibration?
The CYCLERtest calibration can be carried out with either the heated lid being open or closed. Measuring with the heated lid closed can lead to differences, depensing on how the thermocycler reacts on closing the lid. We typically see improvements in uniformity as the thermocycler does not loose heat to the environment and 0.2-0.4C higher accuracies, but for some thermocyclers there is no difference at all. For the Mx3000/3005 you see a deterioration if the uniformity.

6. How often do METTLER TOLEDO advise a thermal cycler should be calibrated?
We advise to have a calibration test at least every year, or every 250 runs, whatever comes first. Some laboratory, national or international regulations might require you to do this more often. We would therefore suggest you take advice from your accredited authority.

7. Isn’t it possible to do a temperature measurement myself?
It will be very difficult. CYCLERtest uses the MTAS (Mobile Temperarure Acquisition System) that was developed by GENOtronics. It uses very sophisticated temperature sensors (thermistors), electronics and software algorithms, to correct for possible deviations. The overall accuracy of this system is better than 0.1ºC. It also makes dynamic multichannel measurements.

8. What does thermal cycler calibration mean and what benefit is it to me?
We test your thermal cycler against internationally accepted standards, and the measurement system itself is complying with the offical temperature measurements regulations. This means that if you have your thermal cycler performance tested with our system, it is also calibrated according to these regulations.

9. What if I buy a new model thermal cycler - how can I obtain the specification sets for my Driftcon system?
You can purchase these from METTLER TOLEDO. The specifications will be loaded onto your computer when you synchronise with CYCLERtest via the Driftcon software.

10. What if I want to check my cyclers’ performance more often?
We have also developed the Driftcon system for checking the performance of your cycler on a monthly, weekly or daily basis. It is an end user temperature measurement fixture that will show the drift of the accuracy and the spread of your thermal cycler in time, and in relation with the global field specifications of that brand and type of cycler.

11. What if my machine fails after a CYCLERtest calibration?
If your machine fails, the generated report will allow you to make an informed decision based upon the errors found. If the errors are small you may decide they are acceptable. If the errors are found in a specific area then this would suggest a mechanical fault and therefore repairs would be needed. You would then need to return your thermal cycler to the manufacturer.

12. What is an overshoot and an undershoot?
Due to the lack of control, cyclers often are not able to reach a target temperature in a stable way. This means that a denaturation temperature can go much higher that the set temperature, before reaching a plateau. For the annealing temperature plateau, undershoots of up to several degrees are not uncommon.

13. What is the definition of accuracy and spread?
Accuracy is defined as the difference in display and the actual block temperature. Spread (or non-uniformity) is the difference in the temperature between the wells of a cycler block.

14. What is the difference between sending my thermal cycler away to the manufacturer for servicing and METTLER TOLEDO performing the CYCLERtest calibration?
The CYCLERtest (MTAS) calibration is a far more detailed and dynamic calibration check compared to that carried out when your machine is serviced by the manufacturer.

METTLER TOLEDO by using CYCLERtest will provide you with a comprehensive report on how your machine is performing. We will not service the machine and repair problems which are found during the test.

15. What kind of report/documentation do I receive with a CYCLERtest calibration?
The test report is encrypted and so can not be modified by an external source. A certificate of Thermal Calibration accompanies every test report, this certificate represents the results of the measured and calculated parameters of the tested instrument.

16. What’s the warranty on your calibration and performance test?
There is no ’warranty’ on the test. We show the output performance of your cycler at the time of measurement. The performance measurement cannot predict what will happen with the accuracy and spread of your cycler in time, nor will it predict if your cycler breaks down the next day. Thermal cyclers wear in time and so will their performance.

17. Which thermal cycler is the best?
Despite all the data that has been collated, CYCLERtest and METTLER TOLEDO will refrain from making any statements on this issue. First of all, we want to be completely independent. Secondly, it is very difficult to make general statements about certain brands or types, because there are already hugh differences between machines of the same type or brand, and also the age of the cycler is of influence.

18. Why are the probes placed in specific well positions?
The positioning of the probes has been devised by using empirical and mathematical data in order to place the probes in the optimum positions for accurate and precise temperature measurements.

19. Why do METTLER TOLEDO not test all 96 wells of the block?
Each well is not individually controlled by the heating and cooling system therefore the positions selected allow an overview of the block to be ascertained, however if you specifically require all wells to be tested this can be carried out. Please contact METTLER TOLEDO to discuss this further.

20. Why should I test my thermal cycler? My results are always OK...
It is very important to know how your thermal cycler behaves. Negative results could be caused by temperature differences in the block, and could be positive in reality. Biological controls are not sufficient to compensate for these differences as variances can occur between wells.