Bio-Rad
T100™ Thermal Cycler
186-1096
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In the age of digital and real-time PCR, traditional PCR may have been cornered to the backseat. However, for isolation of large amounts of amplified DNA, it is still considered to be cheaper using traditional PCR. Hence, it still finds a place today in most research laboratories. Identification and amplification of viral cDNA was critical for our quantification purposes especially for obtaining large samples of cDNA to be used as standards. Hence, we needed a robust and relatively cheaper thermal block for quick amplification and recovery using conventional electrophoresis.
Amplification of viral cDNA
Isolated viral RNA reverse transcribed to cDNA
Following is a summary of the PCR part of the protocol used. Total Volume: 100 ulPCR Mix: 90 ulForward Primer: 400 nMReverse Primer: 400 nMcDNA Sample: 6 ulIncubation: 50 deg C for 2 mins Activation: 95 deg C for 10 mins PCR cycles: 40 Denaturation step in each cycle: 95 deg C for 15 sec Annealing and Extension in each cycle: 60 deg C for 1 min
Make sure the sample block is sealed well
As shown in the image, the amplification of 2 our viral RNA samples was very efficient and a gel run showed the presence of only 1 band in each lane (2 and 3; 1 is ladder)
Quick, saves a lot of protocols for easy recall
None
Quick and Easy to setup.
None so far
Traditional PCR still has a place in most life science laboratories and this product delivers quality results.
Product Name: T100™ Thermal CyclerSupplier Page