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Many different approaches have been used for identifying known mutations. Usually starting with the polymerase chain reaction (PCR), additional assay steps are performed based on the type of mutation. Table 5 shows examples of some of the frequently used techniques as well as their advantages and disadvantages. Here, a brief view of some of these techniques is presented with focus on their applications.
Polymerase chain reaction (PCR) and its versions: In 1980s, Dr Mullis introduced a method for amplifying DNA fragment to a large number of fragments in only a few hours; this method, named polymerase chain reaction (PCR), was a critical point in molecular biology[29,30]. Essential components of polymerase chain reaction are template DNA, primers (a pair of synthetic oligonucleotides complementary to the two strands of target DNA), thermostable DNA polymerase enzyme (e.g. Taq), divalent cations (usually Mg2+), deoxynucleoside triphosphates (dNTPs) and buffer solution. PCR, consisting of 25-40 repeated cycles, has three discrete steps of temperature changes (Fig. 3A); after a single temperature step at a high temperature (>90°C), a series of cycles of denaturation, annealing of primers and extension are performed and followed by a single temperature step called final product extension or brief storage. These steps are as follow:
Initial de


Original text

Many different approaches have been used for identifying known mutations. Usually starting with the polymerase chain reaction (PCR), additional assay steps are performed based on the type of mutation. Table 5 shows examples of some of the frequently used techniques as well as their advantages and disadvantages. Here, a brief view of some of these techniques is presented with focus on their applications.
Polymerase chain reaction (PCR) and its versions: In 1980s, Dr Mullis introduced a method for amplifying DNA fragment to a large number of fragments in only a few hours; this method, named polymerase chain reaction (PCR), was a critical point in molecular biology[29,30]. Essential components of polymerase chain reaction are template DNA, primers (a pair of synthetic oligonucleotides complementary to the two strands of target DNA), thermostable DNA polymerase enzyme (e.g. Taq), divalent cations (usually Mg2+), deoxynucleoside triphosphates (dNTPs) and buffer solution. PCR, consisting of 25-40 repeated cycles, has three discrete steps of temperature changes (Fig. 3A); after a single temperature step at a high temperature (>90°C), a series of cycles of denaturation, annealing of primers and extension are performed and followed by a single temperature step called final product extension or brief storage. These steps are as follow:
Initial de

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