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Specific DNA segments defined by the sequence of two oligonucleotides can be enzymatically amplified up to a millionfold using the polymerase chain reaction (PCR). One of the most significant uses of this technique is for generation of sequencing templates, either from cloned inserts or directly from genomic DNA.
Nowadays, the direct sequencing of PCR products has already played a significant role in molecular biology and genomics research. Such sequencing is widely applied to the detection of gene mutation, diagnosis of genetic diseases, and polymorphism research of single nucleotide. Compared with traditional clone sequencing, direct sequencing of PCR products conducts sequencing towards the amplified DNA directly, which eliminates time-consuming cloning procedures and avoids the traditional repetitive operations like extraction of template. In this way, the correct DNA sequence information can be received from a small number of original samples. Direct sequencing of PCR products are equipped with the following advantages: fast, convenient, simple, and stable.