A validated microarray platform for Mycobacterium tuberculosis

The new microarray described for Mycobacterium tuberculosis in our study has a more complete reprensentation of the genome than any other array design reported till date. Further, protocols for sample preparation, labelling and hybridisation for accurate gene expression profiling of M.tuberculosis have been optimised. Whole genome expression profiling on Mycobacterium tuberculosis H37Rv (OD600 0.4-0.5) was performed using exponential phase cultures after 0 and 6 Hrs in presence and absence of drug (Isoniazid) by using PolyA-dT and WT method. The exponential culture after 24 and 72 Hrs were used for validating the specific hybridization with or without formamide. All time points had two biological replicates with two technical replicates.

本研究中针对结核分枝杆菌（Mycobacterium tuberculosis）开发的新型微阵列（microarray），相比迄今已报道的所有阵列设计，其基因组覆盖完整性更优。进一步，本研究优化了针对结核分枝杆菌的样品制备、标记及杂交实验流程，可实现精准的基因表达谱分析。本研究以OD₆₀₀为0.4~0.5的指数生长期结核分枝杆菌H37Rv菌株培养液为材料，采用PolyA-dT与WT两种方法，分别在添加与不添加异烟肼（Isoniazid）的条件下培养0小时和6小时后，开展全基因组表达谱分析。此外，选取培养24小时与72小时的指数生长期培养液，用于验证添加与不添加甲酰胺时的特异性杂交效果。所有时间点均设置2份生物学重复与2份技术重复。