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real time PCR


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7 replies to this topic

#1 lula

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Posted 03 June 2010 - 11:50 PM

hi everty body
im about to buy a REAL time PCR device so can any one tell me what the specifications i should look for and what are the best companies
thank u alll

#2 tea-test

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Posted 04 June 2010 - 09:01 AM

Hi, I can recommend instruments from Applied Biosystems. I would say, they are the leading company in this field because they are not just offering the hardware but also have a large supply with reagents and real time PCR assays available including the low density array technology, although a bit pricey. Also take into account how much support the company can offer, when I have a question I'm really talking to qPCR specialists at ABI, this should be not underestimated. there is also a very powerful qPCR analysis program for ABI instruments available -> real time StatMiner, which can be also linked to Ingenuity to go for the whole gene pathway analysis. I'm not using this software, I just attended a webinar once and it was amazing.
so this is definitely more than just analyzing Ct values. I don't know if it works with other instruments but I don't think so.

in order to give you some recommendations you should give us first some informations what you intend to do, like are 48 wells enough or do you need the 1536 wells?

Edited by tea-test, 04 June 2010 - 09:03 AM.

tea-test: The artist formerly known as Ned Land

#3 lula

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Posted 04 June 2010 - 12:43 PM

hi
thanks for replying
i guess 48 wells are quit enough


















Hi, I can recommend instruments from Applied Biosystems. I would say, they are the leading company in this field because they are not just offering the hardware but also have a large supply with reagents and real time PCR assays available including the low density array technology, although a bit pricey. Also take into account how much support the company can offer, when I have a question I'm really talking to qPCR specialists at ABI, this should be not underestimated. there is also a very powerful qPCR analysis program for ABI instruments available -> real time StatMiner, which can be also linked to Ingenuity to go for the whole gene pathway analysis. I'm not using this software, I just attended a webinar once and it was amazing.
so this is definitely more than just analyzing Ct values. I don't know if it works with other instruments but I don't think so.

in order to give you some recommendations you should give us first some informations what you intend to do, like are 48 wells enough or do you need the 1536 wells?



#4 Trof

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Posted 07 June 2010 - 04:35 AM

Roche or RotorGene make good cyclers too, but it will depend on what aplications are you planning to do with the cycler, and what format you prefer.

If you really put up with 48 wells then the StepOne system from Applied Biosystems is probably the best and cheapest choice. But you have to realize, that if you run for example a standard curve on a run and triplicate for each sample, 48 usually isn't enough. Anyway there is a StepOne Plus for 96 wells to.

RotorGene and Roche LightCycler use capillaries instead of tubes, which can be less comfortable to handle, RotorGene on the other hand offers the best high resolution melting there is. If you plan to use HRM in the future you should consider it. LightCycler 480 is a plate-based and can do HRM too.

If you plan to use ready-to-use real-time kits, you should look in the manual which cycler they recomend.

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#5 lula

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Posted 07 June 2010 - 12:29 PM

thanks for the detailed reply
im really so new at the real time whole system as for years my lab was only working in conventional PCR and RT PCR
i dont even know who the device will work its just i have a found so i bought anthos ELISA AND WILL BUY real time pcr and gel documentation system
do u also know whats the best gel documentation system and are there web site sthat can teach u the principles of real time devices and ELISA so that u can work on them for the first time ?

#6 tea-test

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Posted 09 June 2010 - 08:22 AM

48 wells are really not very much and you are restricted to 48-well plates which is also not a very common format. so, unless you are sure that you will process only a very small set of samples I would not recommend it.
btw, the roche lightcycler 480 doesn't use capillaries but plates (96 or 384). I think if you choose a 96-well machine you cannot do much wrong in face of throughput. and with the 96-well format you have the possibility to use multichannel pipettes which makes the pipetting much easier and faster.

I don't know if it is of interest for you but there will be a new ABI flagship device launched soon (ViiA 7). Maybe they are then doing a sale of the old ones :wacko:
http://marketing.app...ing?cid=we11913
tea-test: The artist formerly known as Ned Land

#7 lula

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Posted 10 June 2010 - 09:28 AM

THANKS FOR REPLYING
I THINK U R RIGHT ABOUT THE 48 WELL SYSTEM , BUT I HOPE MY BUDGET WILL COVER

#8 tea-test

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Posted 11 June 2010 - 02:54 AM

hi, it is also possible to upgrade the 48well ABI machine later to 96wells, if I understood correct. so you may start with the smaller system and look if you will need the possibility of higher sample throuput.
tea-test: The artist formerly known as Ned Land




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