Assessment of the quality of dna extracted by two techniques from Mycobacterium tuberculosis for fast molecular identification and genotyping

Abstract

We report a comparative study of two DNA extraction techniques, thermolysis and chemical lysis (CTAB), for molecular identification and genotyping of M. tuberculosis. Forty DNA samples were subjected to PCR and the results demonstrated that with thermolysis it is possible to obtain useful data that enables fast identification and genotyping

Parallel susceptibility testing of bacteria through culture-quantitative PCR in 96-well plates

“…The thermo-cold lysis method was modified from that reported by Miyata et al (2011) PCR 8-tube strip was centrifuged at 2000 g for 5 min and the supernatants were ready to be used as the DNA templates for the qPCR detection. For comparison, the bacterial genomic DNA was also extracted and purified from the suspensions using a commercial bacterial extract kit (Omega Bacterial DNA Kit) based on silica-column principle and direct PCR without any extraction using bacteria culture.…”

Initial experiences withMycobacterium tuberculosisDNA extraction for downstream Deeplex Myc-TB targeted deep sequencing in a high burden setting

“…tuberculosis is known for its unique cell wall, which is tough and contains high levels of mycolic acids and other lipids that make it difficult to lyse. There is substantial variation in the published lysis (e.g., sonication, chemical, heat, and bead-beating) and DNA extraction methods (e.g., phenol-chloroform extraction, ethanol precipitation, and column and bead-based methods) for M. tuberculosis, which result in differences in DNA yield, purity, and quality [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17] . In addition, the same method is seldom used by more than one group, and the measure of method success varies.…”