Analysis of TPOX short tandem repeat locus with matrix-associated laser desorption/ionization time-of-flight-based restriction fragment mass polymorphism assay.
Short tandem repeat (STR) loci are routinely analyzed by capillary electrophoresis. However, this method has several disadvantages, including long operational time, low throughput, and inaccuracy. As a result of the introduction of matrix-associated laser desorption/ionization time-of-flight (MALDI-TOF) and electrospray ionization (ESI), mass spectrometry has become an alternative method for genotyping polymorphic STR loci. Here we established a restriction fragment mass polymorphism (RFMP) assay for genotyping STR locus, TPOX, by typeIIS restriction endonuclease cleavage of polymerase chain reaction (PCR) amplicon followed by MALDI-TOF mass spectrometry. The resulting TPOX genotypes from this assay were in good agreement with the results from direct DNA sequencing and GeneScan assays. Our results showed that the RFMP assay is an accurate and high-throughput method for analyzing long DNA fragments such as STR markers. Further research with multiple STR loci may allow this assay to be used for diverse applications such as forensics, paternity tests, and detection of genetic disorders.
Research and Development Center, GeneMatrix, Yongin-si, Gyeonggi-do 449-913, Republic of Korea.
Yoon Seok
YS
Cha
Suh Hee
SH
Choi
Joo-Hyoung
JH
Lee
Soo-Kyung
SK
Shin
Seung Hwan
SH
Lee
Soong Deok
SD
Lee
Soo-Ok
SO
Kim
Sun Pyo
SP
Hong
D000483:Alleles; D056426:Genetic Loci; D005838:Genotype; D006801:Humans; D007453:Iodide Peroxidase / Q000235:genetics*; D018895:Microsatellite Repeats; D012150:Polymorphism, Restriction Fragment Length; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D007453:Iodide Peroxidase