Preparation and optimization of gel-based DNA microarray for NAT2 genotyping

Aim To establish a 3-D polyacrylamide gel-based microarray method to detect genotypes of N-acetyltransferase2(NAT2)of patients.Methods The important single nucleotide polymorphisms(SNPs)sequences of NAT2 were searched.Primers and probes were designed and synthesized aided by computer.NAT2 DNA microarray was prepared by dotting the products of PCR,polymerized by TEMED vapor.The products gained by PCR and hybridization were scanned and analyzed.Results NAT2 DNA microarray was prepared for the detection of SNPs.The optimization of PCR and hybridization were executed successfully.At the same time,constituted criteria for classification were constructed successfully.The signal intensity ratio of Cy3 to Cy5 for the wild type homozygote is ≥5,and mutant is ≤0.2,when the ratio is between 0.6~1.5,it is heterozygote.The genotyping results of 20 samples were tested by direct sequencing,all genotyping results detected by microarray conformed to the direct sequencing.Conclusion NAT2 DNA microarray demonstrated that this is a reliable novel method for NAT2 genotyping;it can be used in rational drug therapy.