Apolipoprotein E (APOE) plays a critical role in lipoprotein metabolism by binding to both low-density lipoprotein and APOE receptors. The APOE gene has three allelic forms, ε2, ε3, and ε4, which encode different isoforms of the APOE protein. In this study, we have developed a new genotyping method for APOE. Our multiplex tetra-primer amplification refractory mutation system (multiplex T-ARMS) polymerase chain reaction (PCR) was performed in a single reaction tube with six primers consisting of two common primers and two specific primers for each of two single nucleotide polymorphism (SNP) sites. We obtained definitive electropherograms that showed three (ε2/ε2, ε3/ε3, and ε4/ε4), four (ε2/ε3 and ε3/ε4), and five (ε2/ε4) amplicons by multiplex T-ARMS PCR in a single reaction tube. Multiplex T-ARMS PCR for APOE genotyping is a simple and accurate method that requires only a single PCR reaction, without any another treatments or expensive instrumentation, to simultaneously identify two sites of single nucleotide polymorphisms.
Bibliographical noteFunding Information:
This work was supported by the Korea Science and Engineering Foundation (KOSEF) through the National Research Lab. Program (No. M1040000029706J000029710) and the National R&D program (M10640010002-06N4001-00210) funded by the Ministry of Science and Technology (MOST).
- Amplification refractory mutation system
- Apolipoprotein E
- Polymerase chain reaction
- Single nucleotide polymorphism
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology