ABSTRACT
A typical thermal cycling protocol is given in Table 4.2. After an initial denaturation step at 95°C, steps 2, 3, and 4 are then sequentially performed for 35 cycles, followed by step 5 at 72°C to ensure that the final round of strand synthesis at high substrate concentration is completed. The 6th step, involving reduction of the temperature to 4°C, is used to terminate all reactions for convenient holding until agarose gel electrophoresis is performed. The time required to traverse from one temperature to another is referred to as the
Figure 4.1 Amplification of a known target sequence with a set of two primers.
