Which amplification method would NOT typically be considered isothermal?

Real-time PCR, which is the method identified as not typically being isothermal, relies on thermal cycling to achieve its amplification. This method operates by repeatedly denaturing the DNA at a high temperature, allowing for the annealing of primers at a lower temperature, followed by extension. These temperature transitions are essential for the PCR process to function effectively, as they facilitate the necessary biochemical reactions for amplification.

In contrast, the other methods mentioned maintain a constant temperature throughout the process. Strand Displacement Amplification, Loop-mediated isothermal amplification (LAMP), and Transcription-mediated amplification system (TAS) all function at isothermal conditions, meaning they do not require the temperature cycling that is characteristic of conventional PCR. This makes these methods advantageous in certain applications where thermal cycling can be a limitation, such as in portable or field-based testing scenarios.