The comparison of different wall-breaking method for extracting total RNA of Mycobacterium tuberculosis

Objective To establish a simple and effective total RNA extraction method of Mycobacterium tuberculosis.Methods Cell wall of Mycobacterium tuberculosis were treated by ultrasonic and glass-bead associated with Trizol method respectively,the total RNA was extracted and the influence of different total RNA extraction methods of Mtb were evaluated through fluorescent quantitative RT-PCR.The negative control is ultrapure water instead of template.Each method were extracted from 3 samples for testing.The final result is the average of 3 specimen.Results The Ct and △Rn value of glass-bead group is 20.67 and 0.644,400 W ultrasonic group is 22.09 and 0.571,240 W ultrasonic group is 21.86 and 0.503,160 W ultrasonic group is 25.21 and 0.411,bead absent group is 28.40 and 0.299,negative control is 40.00 and 0.000.Conclusion The wall-breaking effection of glass-bead method is better than that of ultrasonic method.