PCR-product microarray based on polyacrylic acid-modified surface for SNP genotyping.
PCR-product microarray has great efficiency in SNP genotyping, mutation screening and epigenetic analyzing from a large number of samples. The current PCR-product microarray technology is mostly based on the 3-D gel microarray technologies due to its high loading capacity for PCR products, while there is little progress for PCR-product microarray on planar glass, which gives low background and convenient fabrication. In this study, we improved the PCR-product microarray on planar glass by employing a polyacrylic acid-covered slide. The raw amino-modified PCR products were simply precipitated with ethanol and directly spotted for DNA immobilization. Three detection methods of hybridization, solid-single base extension and solid-multiple bases elongation were carried out for single nucleotide variation identification on the PCR-product microarray. The experimental results showed that the high immobilization yield for raw PCR product was achieved, and the high specificity and high ratio of S/N for genotyping on the microarray were obtained. SNP genotyping of cytochrome P450 2D6 gene in 30 individuals was successfully demonstrated. This study has significantly increased the performances of PCR-product microarray, which could improve its applications in SNP genotyping and mutation screening in large number of individuals.
Shi
Xiaolong
X
State Key Laboratory of Bioelectronics, Southeast University, Nanjing, PR China.
Tang
Chao
C
Zhou
Dequan
D
Sun
Jinxia
J
Lu
Zuhong
Z
D000180:Acrylic Resins / Q000737:chemistry*; D019389:Cytochrome P-450 CYP2D6 / Q000235:genetics; D005453:Fluorescence; D005838:Genotype; D006801:Humans; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012680:Sensitivity and Specificity
D000180:Acrylic Resins; C006903:carbopol 940; D019389:Cytochrome P-450 CYP2D6