I am looking for a PCR machine for my lab. Which specifications are most important? And what does it mean when the thermal is Peltier-based designed?
As others said:
1-Block capacity
2- the tube volume that fit well in PCR block
3- Gradient module
4- Ramp Temp and time
5- Number of Block
6- User interface
7- Its Price
Almost all newly manufactured PCR machine comes with Hot-lid, so it is not a matter of choice in selecting a PCR machine.
Important:
1. Bottom and top lid heated.
2. Temperature gradient mode.
Otherwise all the cyclers seem to be similar.
Peltier cooling is a mechanism of cooling which is purely solid-state, and is basically quieter and more accurate than traditional radiator based systems. Basically its more accurate and most likely faster.
If you work with animals and need to do mass genotyping, and plan on having a large animal colony, a PCR machine with 96 wells helps greatly, as does a dual head, which allows two different PCR programs to run.
So those are more the physical aspects which make it useful if you plan on doing lots of PCR, primarily genotyping.
In terms of numerical specs, the faster it is able to ramp up and down temperatures will drastically reduce the time needed for PCRs. We have an old pcr machine, where a touchdown program requires 4 hours to run, while a newer one only needs 2 hours, purely because of its speed in changing temperatures.
Hope you find a good PCR machine for your lab!
Peltier based design is how the block is heated/cooled. The peltier effect is a thermoelectric phenomenon by which materials can be heated/cooled rapidly. This rapid ramping is desirable when doing pcr. These days, its almost a given that a pcr machine will use the peltier effect to achieve desired temperatures.
In terms of specifications, you ideally want something with a fast temperature ramp up/cool down time, and with low error. Thats basically it when it comes to pcr machines. Its nothing more than a machine which changes temperatures.
Depending upon your budget, you can get additional bells and whistles in the machine, such as..
1. Number of pcr tubes it can hold
2. Size of the wells...depending upon the pcr volume.
3. Number of users: if there are going to be multiple users, there are machines which can run 2 independent pcr's at the same time.
4. If you are going to be doing a lot of pcr's for which you need to fine tune annealing temp for different primers, a gradient pcr machine is desirable....allows you to run multiple different annealing temps in the same run, thus saving time.
These things can help you determine what would be a good choice for a pcr machine.
Ramping time (how fast the block cools and heats) and temperature uniformity among the wells are probably the two most important. Peltier-based means refers to the 'Peltier effect', it uses a current applied across a metal to raise and lower temps very rapidly. I would suggest getting a cycler with a gradient function if you can, these always come in very handy.
We have MJ Research PCR machines which do the gradient as well as normal PCRs. I have heard about the Thermo Fischer PCR ones too where we can log all our work. Roche real-time PCR machines are user-friendly. I think the most important specifications would be to use both gradient and the normal PCR and also to have something where in we can keep a log of our work. But if you need a real-time one, then it is Roche.
Eppendorf Mastercycler gradient cycler is a good one.having the all the qualities that has been described by others.
I have used three PCR machines. Bio-Rad DNA Engine is the best one which does gradient and has fast ramping time.
Besides that the technical aspects of the PCR machine should meet your requirements, regular PCR or real-time, which will very much depend on your applications and budget, do not forget to take the software into account. This is especially important for real-time PCR machines, which can range in price from E 30.000 [eg Agilent/Stratagene's MX3000] to up to E 100.000 [Roche/Applied Biosystems].
For a real-time PCR machine, which can do regular PCRs as well, you would like to have a window-based software package that can be installed on any PC without having to buy additional licenses and that does get updated regularly taking new applications into consideration.
with my experience with several thermocyclers, i will readily recommend Biometra T professional, it a very good ramping time,and rubost.
As others said:
1-Block capacity
2- the tube volume that fit well in PCR block
3- Gradient module
4- Ramp Temp and time
5- Number of Block
6- User interface
7- Its Price
Almost all newly manufactured PCR machine comes with Hot-lid, so it is not a matter of choice in selecting a PCR machine.
Dear Alethea,
We have used the MJ research Thermal (Peltier-based and hot lid) for many years and it is very good. We have three in the lab, with and without gradient. This machine also has an affordable price. I strongly recommend.
Good luck !!!
Claudete
Univ. Londrina, PR
Obrigada Claudete, pela sugestão. Eu já trabalhei com a marca MJ Research, e gostei muito. Aqui no Ceara, eu só vi maquinas da Applied, Eppendorf e Bio-Rad. Com exceção da Applied, todas as outras já deram problemas. Também gostei muito do software da MJ, fácil de usar e prático. Vou seguir sua recomendação. Obrigada novamente. Aletheia.
I have used both eppendorf and VERTI of Applied biosystem....
1. the ramp speed,
2. wide temperature range
3. gradient temp (we can set what we want - not like eppendorf gradient)
4. 96 wells
5. pause option
6. price range
7. services
In all these aspect is strongly suggest VERTI of Applied biosystem now it is call as "Life technologies".
One important aspect I think left is for having constant distribution of temperature of hot lid over the tubes. Lid made of solid metal may not put equal pressure and temperature that causes evaporation of reaction mix from corner/border placed tubed. As suggested by Meenakshi, Eppendorf PCR has this facility with cushion lids . So do consider provision of this facility also.
no. of blocks
no. of wells
ramping rate
gradient module
pause and auto restart option in case of power failure
material of the block, aluminium blocks are the best for uniform temperature distribution
no. of peltiers (more is better)
tempreature uniformity and accuracy (ideally between 0.1 degree C and 0.25 degree C)
Hi,
In our lab, we have two of these: 2720 Thermal Cycler; from http://www.appliedbiosystems.com/absite/us/en/home/applications-technologies/pcr/thermal-cyclers.html
We are very satisfied with them.
Good luck.
Marisa
Institute for Adriatic Crops
Croatia
I recommend geneAmp 9700 applied biosystem very good and very robust
Good luck
Biometra's thermal cyclers work really well. We have older cyclers, 10 years old and they are still in good condition like newest ones, also from Biometra.
Hi Aletheia,
You're right, there are so many machines.
To find the right one, it depends of the money you have to buy it, and the use of the machine.
- one important option is to have the hot lid, or else you will have to add mineral oil in all your tubes before starting th PCR.
- second, if you have to optimize PCR conditions and to test different annealing temperature in one PCR, the temperature gradient option is interesting, but is expensive.
- third the ramp speed is important for good PCR results (betwenn 3 and 6°C/sec).
- If you want to reduce PCR time, choose a Silver block instead of an aluminium block (better temperature conducer).
- pause option is interesting in some protocols, or to maintain a low temperature during the night if you start your PCR at the end of the day.
My preferred machine are from MJ research and Eppendorf.
I also had bad experience with BioRad and Techne.
Tchao bella
Peltier define the technology used to heat and cool the samples, at present almost all the PCR thermal cycler are peltier based. Important specification are (not necessarily in the order)
1) Ramp speed
2) Temperature range
3) Gradient option
4) Price
F) Format (compatibility with different plates brand)
6) Service in term of custorer service (really critical)
For me important specifications are
1. Temperature range
2. Gradient option
3. No. Of wells
4. Equal distribution of temperature over multiple samples
5. Price
6. Service
See that the repair service is near. Otherwise, PCR machine should be fast and there are big differences in that.
On the other hand: if thermal cycler needs service, should be rejected from list of candidate stuff. :P
BTW choice of parameters depends also from you and your expectations: if you want to do PCR's in the same conditions as some described previously in some papers and they are really diffcult, it's better to look for some similar cyclers with similar parameters. You exclude possibility that something will fail and you will have to find a reason looking for optimisation, DNA, and many other reasons of lack of results.
I think the following online material is helpful in this regard
http://www.conflutech.net/_data/PCR_5_Lab_setup_.pdf
Prezada Aletheia. As dicas que vc recebeu são fundamentais e deverão ser levadas em consideração sempre! Te sugiro também te preocupares com assistência técnica, procure verificar se a empresa que comercializará o equipamento poderá substituir a sua máquina caso ela apresente defito. Isso garantirá a manutenção das suas pesquisas. Um bom termociclador é essencial para um lab de biomol. Sds.
Obrigada Nelson, pelas dicas. Eu vou levar tudo em consideração, porque a maquina da Biorad está parada a mais de 01 mês por conta da manutenção dificil...
When I was trying to find a new PCR machine for my lab, we also looked at temperature accuracy. Some machines overshoot or undershoot the annealing temperature, so they are not as accurate to the programmed temperature. There are several papers that compare different PCR machines and different technical aspects.
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