Thanks to all those who responded to my question about which real-time PCR machine to purchase. What follows is a summary of the responses I got. Our company has used 3 of the 4 you mentioned: ABI 7300 BioRad DNA Engine Opticon 2 (this was also MJR) BioRad MyiQ And we've even tested out the Stratagene MX3005P It all comes down to what special preferences you would want from the machines. ABI: We've had some limitations in programming options with this machine, however, we're not doing the typical real-time PCR assays. Opticon2/Stratagene: Very user friendly. Of these two, we prefer the Opticon2 because of the nice features in viewing your traces (you can roll over the trace and it will highlight in different colors which sample it is both on the plate position as well as in the description - VERY useful if running tons of samples at the same time). MyiQ: A busy & slightly complicated interface (lots of tabs and buttons to activate before you can edit/create anything) but after some practice, it isn't too difficult. Does not have the roll-over-trace feature; instead, you have to select the wells you want display or place your cursor over the trace and catch the little hand cursor for it to tell you which sample the trace is belongs to (a bit annoying when looking at several samples, but BioRad is working on the feature for their next softward upgrade'at least I hope). Bonus feature is that you can add cycles /during/ a run. The Opticon2 is limited to 99 cycles (again, the types of assays we're doing requires us to sometimes run the whole thing out for a long time to see when a product will form). It also has a much faster ramp time then the Opticon2. Something that is important to us is the ability to pause the run and add/remove small amounts of volume in our sample and both of the BioRad machines were the only ones that allowed us to do that (meaning the plate/tubes were not physically being blocked by the laser or whatnot when we interrupt a run). We currently own all three (ABI and both BioRads) and will probably consider buying another MyiQ. _____________ you might look into the stratagene machine. we have one and love it. + ___________ ______________ Hello, Alex, I'll forward to you my reply to Cort, who sent in a similar query to the evoldir not long ago. Best of luck, Bob PS: Note that the Eppendorf is pretty high priced (around same as ABI 7500, I think). I'd stick with the Bio-Rad rather than the MJ, because I think you'll get better customer support on the iQ series. --------- *From:* Robert E. Marra [mailto:Robert.Marra@po.state.ct.us] *Sent:* Wednesday, October 11, 2006 8:36 AM *To:* 'cla@uidaho.edu' *Subject:* realtime decisions Hello Cort, I shopped around for a realtime instrument a little over a year ago. Although there seems to be a penchant for the Cepheid Smartcycler in my field, for diagnostics, I dismissed it because of its sample size limitation (16 per run). I looked at the ABIs, the Bio-Rad iQ5, and the Eppendorf. I briefly looked into the Stratagene, but did not like several features, including the moving camera, and I was nervous about its low price, wondering if it wouldn't come back to haunt me; i.e., you get what you pay for. I only looked at the iQ5 from Bio-Rad, not the MJResearch, and that was in part because at the time, Bio-Rad was in litigation with ABI over licensing for their MJR line (perhaps you know that Bio-Rad purchased MJR some time ago). We (my technician and I) were interested in the software as well as the instrument itself. One thing that appealed to us about the iQ5 and the Eppendorf is that both instruments can be used as stand-alone thermalcyclers, so part of my purchasing was to buy one real-time unit and two stand-alone thermalcyclers, and I liked the idea that all three would share the same base unit, so that reaction optimization can be done any machine, and for end-point real-time PCR, the reactions themselves can be done in a different unit, than transferred to the iQ5 to be read. The ABI units don't have this feature. I liked the Eppendorf best of all, both the instrument and the software, but there were holdups relating to the licensing litigation, and I had to make my purchase before the opportunity to use the money would run out. So I went with the iQ5, and for the most part, we've been very happy with our decision. For one thing, Eppendorf was not very flexible on the pricing, even considering that we were going to by the realtime unit as well as two thermalcyclers, whereas Bio-Rad worked out a nice deal for us. As with anything, since we've been using the iQ5 (and we use it a lot), we have found some quirks in the software, but they seem to be pretty responsive to suggestions for updates, and they've already incorporated a couple of our suggestions (which, apparently, others made as well). The tech support has been excellent, too. One of my fears with ABI is that while they put a lot of effort into the sale, when it comes time for tech support, they are hard to get hold of. _________________ Dear Alex, some time ago I posted in evoldir a similar question althogh not so selective on models. I am listing the answer I got and I hope you find them useful. Eppendorf thermocycler use capillaries and the software is more complicated to use. Best wishes, Sergio ____ Sergio Stefanni, PhD Dept. of Oceanography and Fisheries (DOP) IMAR-University of the Azores PT-9901-862 Horta - Azores - Portugal Tel: +351.292 200 439 Fax: +351.292 200 411 email: sstefanni@notes.horta.uac.pt / sstefanni@yahoo.co.uk Feedbacks from EvolDir members on RT PCR Thermocyclers: Dear Sergio Stefanni, At the following of your mail, I think I can give you a advice. Since I'm working for the Invitrogen company, I'm aware about qPCR products and apparatus. I have to say that the most common brand we can see is the Applied Biosystem machines. It depends the frequency of yous pCR. If you want to do a lot of them (three to four a day) the Applied 7000 or 7500 is ok, but there is also the new Roche that is the C480 that allow you to perform a lot. But, in term of products you have to use on the machine, the ABI is better because it exist a larger range of products of different brands you can use on these machines and allow you comparisons between the kits and products. If you want to use only SybrGreen for detection, the ABI5000 is better because allows you to use less Rox, that is not so bad, you can save lot of money at the end. If you want to use primers as Taq man technology or the Lux Primers, all of ABI and C480 are ok. You have just to know that the light cycler from Roche is more difficult to use as this is working with capilaries and this is the only machine that exists with capilaries. So, hope it will helps, if you have any questions, you can ask me, no problem, Corinne. Corinne Labrosse, PhD Invitrogen Dear Sergio, Perhaps I wasn't clear... The only capillaries machine is the Roche Ligth Cycler. I would say this is more a disadvantage than a advantage because you have few kits working on it and it oblige to use BSA to avoid material attaching the capillaries. The performances don't' seem better than an other machine. The C480 is a big beat I can say. This machine is usually for people that are using this machine a lot and dooing a lot of reactions a day. This is a plate machine, not a capillaries one. I don't know really well the Bio Rad machine iCycler. The only thing with this one is that the passive reference of fluorecence is Fluoresceine and not the Rox, that is more used for the other machines. Then you will have less choice in terms of kits. After this is as you want. As I told you, the ABI are the most common machines and the 7500 allow you to use less Rox quantities so as you can save money, that is not so bad. Hope this will help, Regards, Corinne. _____ Have a look at Biorad. Cheaper than ABI, work in an easier way, and don't limit you to their own consumables. Also, you can get a representative to come and give you a demo. Amber Teacher Institute of Zoology Zoological Society of London Regent's Park, London NW1 4RY U.K. _____ Hi, my name is Delphine and I work in the university of Rimouski, Québec (Canada). I have worked with three real-time PCR systems: Smartcycler from Cepheid, with a ABI one and with a Biorad one (the IQ one). I do not recommand you to buy a smartcycler. Not because it's not good....it's a really good machine to do optimization but not to do routine experiments. There is only 16 cells in a block (you can run 6 blocks in a same time but each block are really expensive), tubes are really specific (you can only buy their cepheid tubes) and minimum volume is 25ul (you can't diminish it since of the form of the tubes). I like to work with the ABI one and the Biorad one. Both are really good machines. Maybe the Biorad one is less expensive than the ABI. With the ABI one I just used ABI furnitures (plates, products...) but I don't know if other furnitures can do. With the Biorad one I used without any problem tubes or plates or products from others companies. I really like the Biorad one. It does not take a lot of space and it's really easy to use. Biorad offers a smaller machine (the MiQ orsomthing like that). This one is really less expensive and as good as the IQcycler BUT the MiQ as only on channel of detection (for sybr green) whereas the IQcycler as 4 channels of detection. Whatever your choice with the company I recommand you to buy one with four detection channels since you will be able to perform multiplexing. If you want to perform routinely a lot of measures I don't think that you can have a 384 wells block with Biorad (I'm not sure) but you can have it with the ABI one and it really diminish the volume of reaction products. this is what I can tell about my experiences. I hope it will be helpfull. Have a nice day, Delphine _____ Dear Sergio, I bought a Bio-Rad iQ5 last year, and for the most part we are very happy with it. I like the idea that the base unit is identical to the base unit of the standard iCyclers, of which we have two (bought at the same time). There are a few quirks in the software that you never realize until you're actually using it, but the good thing is that Bio-Rad is fairly responsive about incorporating changes in future software upgrades. I didn't want the ABI because of the cost and also because for that amount of money it realy should be a gradient machine (the Bio-Rad and Eppendorf both are gradients, which is actually quite helpful when you're optimizing, no matter what anyone says about doing all your reactions at the same annealing temperature). I really liked the Eppendorf, but at the time I had to make my purchase, Eppendorf was still in arbitration with ABI over licensing rights. They kept promising a release date, only to move it back yet another month, and I couldn't wait any longer. It struck me as very well designed software and like all their other equipment, very well made. I would avoid the Stratagene if I were you. I don't know anything about the MJ units that Bio-Rad now sells. Hope that helps. Bob Robert E. Marra, Ph.D. Plant Pathology & Ecology The Connecticut Agricultural Experiment Station 123 Huntington St New Haven CT 06511 _____ Hi, We have a BioRad iCycler for two years by now. Happy with it. BioRad also has a one color iCycler at a much lower price. Ciao! Xiao-Ru Xiao-Ru Wang, PhD National Institute for Working Life SE-907 13 Umeå, Sweden _____ Dear Dr. Stefanni, In response to your Evoldir posting, I have been used the ABI instruments extensively, and have been using a Stratagene MX3000P for the last 3 years. I strongly recommend the Stratagene instrument, which is inexpensive, reliable, and the software has been much more easy to use than the ABI software. I have also found Stratagene technical help to be very responsive and helpful when I have had problems or questions. I hope this helps. Phil Phillip A. Morin, Ph.D. Molecular Ecology Lab, Protected Resources Division Southwest Fisheries Science Center 8604 La Jolla Shores Drive La Jolla, CA 92037, USA Phone: 858-546-7165 Fax: 858-546-7003 Email: Phillip.Morin@noaa.gov _____ Hi, We have a BioRad iCycler for two years by now. Happy with it. BioRad also has a one color iCycler at a much lower price. Ciao! Xiao-Ru Xiao-Ru Wang, PhD National Institute for Working Life SE-907 13 Umeå, Sweden Phone: +46-90-176 115, Fax: +46-90-176 123 _____ Dear Dr Wilson, An what about the new ligth cycler from Roche : this is probably the most reliable heating block on the market with that of Eppendorf. Unfortunately, all the software are not fully developped yet for the SNP genotyping module, but for the rest, it's OK. We have tested it here with people developping protocols and reagents for qPCR and it's really a good brand. Sincerelly Dr Erick DESMARAIS _____ Hey Alex, You came to the right guy for this one, I have used many many realtime instruments and they are not all created equal, so here is a list of ones I have used onhd my opinion of them: ABI 7300 - all in all a pretty good instrument, has everything you need and is right in the middle price wise (we got ours for a little over 20K, list is about 37K I think). I personally like using this software the best, its user friendly and not overly complicated. Uses 96-well plates and works very well with ABI and BioRad reagents. sensitivity isn't quite as nice as the more expensive ones, and a expression assay takes about 2.5 hrs to run. (don't let them talk you into the fast ramping block ugrade, it's a piece of shit) ABI 7900 - We have this also and use is for very high-throughput projects, it only runs 384 well-plates and is only really practical if you need A LOT of capacity. BioRad iCycler - This one is also an all-around good instrument, the software is a little more confusing than ABI but still has everything you would need and works well with the BioRad reagents wich are a little cheaper. This one is also pretty affordable, we got a good deal because they were pushing them pretty hard. This also runs 96-well plates but like the ABI it does lack a little bit of the sensitivity. this one and the ABI 7300 are pretty much the same in my opinion. Roche Light-cycler - I personally hate this instrument we were a test cite for this instrument for six months and I had to beg people to use it. The worst part about it is that you have to run all the samples in glass capillaries that are hard to load and handle, plus the software is pretty confusing and it's a little pricey. That said this one has great sensitivity and great consistancy in sample data. Eppendorf - This in my opinion is the caddilac of real-time machines, the software is great for set-up and especially analysis, it has the best sensitivity of all the machines I have used, the best consistancy, and it has a lot of really cool features. The down side is its expensive, but if I were you I would price it out, you might be able to get a good deal on one, sometimes they have demo units they will sell you. ____ Assistant Professor Department of Biology University of Miami 1301 Memorial Drive Coral Gables, Florida 33146-0421 USA Phone: (305) 284 2003 Office: Cox Science Room 234 http://www.bio.miami.edu/acwilson/home.htm acwilson@bio.miami.edu