Application of oligonucleotide array for HLA-DQA1 locus genotyping.docVIP

 PAGE \* MERGEFORMAT 11 Application of oligonucleotide array for HLA-DQA1 locus genotyping Author: Wang Tong, Wang Tian-Jiao, He Qun, Yu-Kui Zhang, Jia-Ming Ma, Hou Jian, Wang Shaocheng, Mr Poon, Yu-Jie Zhao 【Abstract】 In order to construct HLA-DQA1 locus and oligonucleotide genotyping chips to take to establish a system used in HLA genotyping integrated technology platform, in the concentration of HLA-DQA1 polymorphism in the second exon region, to design a set of oligonucleotide probe typing; used method of self-built in our laboratory, made oligonucleotide chip. Extraction of genomic DNA with specific primers between the two groups, unilateral primers were fluorescent marker, line asymmetric amplification. Hybridization analysis of hybridization signals detected after the scan to determine the HLA allele; classification results to the standard DNA and PCR products were sequenced and detection. The results showed: 100 cases of the sample chip, genotyping was successful in all 50 cases in which the standard DNA template in line with the other 50 cases of clinical samples randomly selected 10 cases consistent with sequencing results, including two cases of sub-type results are not complete; repeat the hybridization experiments in sites to reproduce the rate of 95%. Conclusion: The development of the HLA-DQA1 allele typing chip, its accuracy and reproducibility of the ideal, and the operation is simple, fast, have extensive application prospects. Keywords: genotyping HLA-DQA1 Genotyping by Using Oligonucleotide Microarrays Abstract In order to fabricate the HLA-DQA1 genotyping chip and develop an integrated, parallel technical platform to type HLA system, a pair of primers and a set of probes were designed according to the sequences of HLA-DQA1 exon 2, where the polymorphism is concentrated. The oligonucleotide chip was made with the methods developed in our laboratory. The target DNA was asymmetrically amplified with the