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PCR consists of a series of 20-40 repeated temperature changes, called thermal cycles, with each cycle commonly consisting of three temperature steps
In the denaturation step, the double-stranded DNA is derctured to obtain single DNA strands by increasing the temperature to 94-98°C for 20-60 seconds
In the following step, the annecling the temperature is lowered to 50-70°C for 30-90 seconds
Those probes are labeled with a donor fluorochrome and a quencher
When the probe is intact, there is no fluorescence
The temperature is increased to 72°C
As the polymerase extends the DNA strands
A final result, the enzyme Teg polymerase synthesizes two new strands of DNA identical to the original strand
The processes of denaturation, annealing and extension constitute a single cycle
After finishing the real time PCR reaction, the PCR product can be detected by measuring the fluorescence produced during the amplification
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