Dna Thermal Cycler
A look at what is currently available on eBay
 PERKIN ELMER DNA THERMAL CYCLER VERSION 2.4 US $125.00 
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 Techne TC-512 Gradient Thermal Cycler PCR DNA Touchscreen LCD Display US $3,499.99
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 Perkin Elmer 480 DNA Thermal Cycler Lab Machine! US $149.99
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 PE Pekin Elmer 480 DNA PCR Thermal Cycler US $219.99
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 PE cetus cycler thermal controller DNA Perkin Elmer US $275.00
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 PERKIN ELMER CETUS DNA THERMAL CYCLER 480 (USED) US $260.00
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 Perkin Elmer Cetus DNA Thermal Cycler Ver 2.2 US $90.00
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 Perkin Elmer PCR DNA Thermal Cycler 480, -5-100°C US $395.00
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 Perkin Elmer GeneAmp PCR DNA Thermal Cycler 9600 US $735.00
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 Perkin Elmer DNA Thermal Cycler 480, 48 well US $130.00
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 Perkin Elmer Cetus DNA Thermal Cycler US $99.00
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 Perkin Elmer Cetus DNA Thermal Thermo Cycler Thermalcycler US $225.00
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 PERKIN ELMER CETUS DNA THERMAL CYCLER 480 -5C TO 100C % US $350.00
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 MJ Research DYAD Disciple PTC-221 DNA Thermal Cycler US $2,950.00
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 Perkin-Elmer Cetus DNA Thermal Cycler 480 US $264.95
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 Biometra UNO-Thermoblock Thermal Cycler PCR DNA US $849.00
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 Ericomp TwinBlock System Easy Cycler series thermal DNA US $899.00
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 PE Perkin Elmer Cetus DNA Thermal Cycler 480 US $199.00
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 Thermolyne Temp Tronic DB66925 DNA Thermal Cycler US $499.00
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 Perkin Elmer 480 DNA Thermal Cycler P/N: N8010100 US $295.00
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 Perkin Elmer 480 DNA Thermal Cycler N801-0110 US $423.20
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How would you do a successful PCR with a GC rich template?
I'm studying genetic engineering, but at a basic level and I can't find the answer to this question in anything but gobbledygook. What do you need to change from the normal PCR? This is the method I have for normal PCR, but I need to find how to do a successful PCR with a DNA fragment that has a high GC fragment.
Move your extracted DNA into a PCR tube, and then add all the materials above, in order.
Place the tube with all the reaction components into a DNA thermal cycler. The first temperature you need is 95oC.
Then cool it down to 50oC.
The next temperature is 72oC
That is one cycle. By the end of cycle three your desired copies will begin to appear.
All I need to know is what needs to be altered in order for a GC rich template to be amplified?
Hi there,
PCR is a funny thing sometimes it works brilliantly straight away and other times not and it can be a bit of trial and error to get it to work.
To amplify a GC rich template I would try adding DMSO or glycerol at a concentration of between 5-10%. This can help improve amplification efficiency and the specificity of the reaction. Adding Betaine to the reaction mix also helps.
It may also help to include an initial denaturing step of 95C usually lasting 2-5 minutes. A PCR template with a high GC content may be hard to denature.
Ensuring that you are using the optimum annealing temperature and time is also important for getting good product amplification. Generally the annealing temperature is related to the GC content of the primer.
Just so there is no confusion with the terms I've used, here is the PCR cycle I usually use.
Initial denaturing step 95C 2-5min
Cycle the following 25 times
Denaturing step 95C 30sec
Annealing step 50-56C 30sec
Elongation Step 72C 1min per 1kb
Final elongation step 72C 5min
Hope this helps.
