PCR efficiency 2026-10-14
For a stipulated population of amplifiable copies, let be the amount after cycle and define PCR efficiency by the fractional new yieldHere means one additional copy per existing copy, hence ideal doubling. If every cycle has the same PCR efficiency , the exponential growth model gives . Starting with copies, twenty ideal cycles give copies; the stipulated value gives about . An efficiency of 80 percent therefore means multiplication by 1.8 each cycle, not retention of 80 percent of the final ideal yield.
In the ideal strand-counting example, all duplex-equivalent material doubles, but exact-length amplicons lag initially. Starting from one longer duplex and retaining its two original strands, cycle contains exact-length strands out of total strands. Thus their fraction is : zero after cycle 1, one quarter after cycle 2 and one half after cycle 3. This counts strands rather than assuming every strand immediately finds an exact-length partner. The generic yield model above ignores this early distinction.
PCR plateau 2026-10-14
A PCR plateau is the late phase in which further cycles add little product. Depletion of substrates and accumulation of reaction by-products can reduce PCR efficiency. Consequently, similar final amplicon amounts need not mean similar initial template amounts. In the stipulated green model, implies , and growth approaches the imposed ceiling. That illustrates loss of starting-amount information without claiming to identify the limiting chemical process in an actual tube.[ref]
PCR Cycling Parameters—Six Key Considerations for Success by Thermo Fisher Scientific. PCR cycle number determination and Figure 7: lowered efficiency and plateau from reaction-component depletion and by-products
Real-time polymerase chain reaction 2026-10-14
Real-time polymerase chain reaction monitors fluorescence during cycling. With validated specificity, calibration and PCR efficiency, that record supports estimating initial template amount; this quantitative use is called qPCR. Ordinary endpoint PCR amplification examines accumulated product afterwards. qPCR need not start from RNA, and RT-PCR need not monitor fluorescence. Combining both operations gives RT-qPCR.[ref]
Real-Time PCR (qPCR) Basics by Thermo Fisher Scientific. Real-Time PCR Basics: amplification monitoring and quantification
RT-qPCR 2026-10-14
RT-qPCR applies qPCR after reverse transcription. Differences in RNA recovery and conversion into Complementary DNA can affect its inferred starting amount as well as differences in PCR efficiency.
