|
|
|
The application of asymmetric PCR-SSCP in gene
mutation detecting |
| ZHANG Xiaohui, XU Shangzhong, GAO Xue, ZHANG Lupei, REN Hongyan, CHEN Jinbao |
| Institute of Animal Sciences, the Chinese Academy of Agricultural Sciences |
|
|
|
|
Abstract The advantages and disadvantages of asymmetric PCR-SSCP and the traditional PCR-SSCP were compared in this study. The mutations in 3′UTR of Smad4 gene of Luxi cattle and the Holstein cow were analyzed by asymmetric PCR-SSCP and one insert “T” mutation and one G/A mutation in this region were found. The G/A mutation created a HhaI restriction enzyme digestion position and the frequencies studied by asymmetric PCR-SSCP and HhaI-RFLP in 116 Luxi cattle and 75 Holstein cows were all the same. The asymmetric PCR-SSCP had fewer, clearer and more stabile bands than traditional PCR-SSCP. This indicates that the asymmetric PCR-SSCP is suited for mutation detection.
|
|
Issue Date: 05 September 2008
|
|
| 1 |
Bhide M R, Mikula I (2005). AsymmetricPCR-SSCP: a useful tool for detection of OLA-DRB1 (MHC Class II) genepolymorphism in Slovak Improved Valachian sheep. Acta Vet Brno, 74: 275–278
|
| 2 |
Boutin P, Hani E H, Vasseur F, Roche C, Bailleul B, Hager J, Froguel P (1997). Automatedfluorescence-based screening for mutation by SSCP: use of universalM13 dye primers for labeling and detection. Biotechniques, 23(3): 358–362
|
| 3 |
Butler J M, Jiang B P, Belgrader P (2001). Automation in genotyping of singlenucleotide polymorphisms. Human Mutation, 17: 475–492.
doi:10.1002/humu.1131
|
| 4 |
Ho-Pun-Cheung A, Choblet S, Colineau T, Abaibou H, Zsoldos D, Brengel-Pesce K, Grenier J, Cleuziat P, Lopez-Crapez E (2006). Detection of single nucleotide polymorphismsby minisequencing on a polypyrrole DNA chip designed for medical diagnosis. Lab Invest, 86(3): 304–313.
doi:10.1038/labinvest.3700387
|
| 5 |
Glavac D, Dean M (1993). Optimizationof the single-strand conformation polymorphism (SSCP) technique fordetection of point mutations. Hum Mutat, 2(5): 404–414.
doi:10.1002/humu.1380020513
|
| 6 |
Isabelle T (1993). Excess PCR primers may dramatically affect SSCP efficiency. Nucleic Acids Research, 21(16): 3909–3910.
doi:10.1093/nar/21.16.3909
|
| 7 |
Kim B C, Park J H, Gu M B (2004). Development of a DNA microarray chipfor the identification of sludge bacteria using an unsequenced randomgenomic DNA hybridization method. EnvironSci Technol, 38(24): 6767–6774.
doi:10.1021/es035398o
|
| 8 |
Kiyama M, Fujita T (1996). High-throughputasymmetric-PCR SSCP analysis using well-controlled temperature conditions. Biotechniques, 21(4): 710–716
|
| 9 |
Kozlowski P, Krzyzosiak W J (2004). Optimumsample medium for single-nucleotide polymorphism and mutation detectionby capillary electrophoresis. Electrophoresis, 25(8): 990–998.
doi:10.1002/elps.200305782
|
| 10 |
Lazaro C, Estivill X (1992). Mutationanalysis of genetic diseases by asymmetric-PCR SSCP and ethidium bromidestaining: application to neurofibromatosis and cystic fibrosis. Mol Cell Probes, 6(5): 357–359.
doi:10.1016/0890‐8508(92)90027‐U
|
| 11 |
Lilleberg S L (2003). In-depth mutation and SNP discovery using DHPLC genescanning. Curr Opin Drug Diseov Devel, 6(2): 237–252
|
| 12 |
Makino R, Kaneko K, Kurahashi T, Matsumura T, Mitamura K (2000). Detectionof mutation of the p53 gene withhigh sensitivity by fluorescence-based PCR-SSCP analysis using low-pHbuffer and an automated DNA sequencer in a large number of DNA samples. Mutation Research, 452(1): 83–90
|
| 13 |
Nishimura A, Tsuhako M (2000). Singlestrand conformation polymorphism analysis of ras oncogene by capillaryelectrophoresis with laser-induced fluorescence detector. Chem Pharm Bull, 48(6): 774–778
|
| 14 |
Ribas G, Neville M J, Campbell R D (2001). Single-nucleotide polymorphism detectionby denaturing high-performance liquid chromatography and direct sequencingin genes in the MHC class III region encoding novel cell surface molecules. Immunogenetics, 53(5): 369–381.
doi:10.1007/s002510100343
|
| 15 |
Vernet G, Tran N (2005). The DNA-Chiptechnology as a new molecular tool for the detection of HBV mutants. Clin Virol, 34(1): 49–53.
doi:10.1016/S1386‐6532(05)80010‐1
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
