Concentration and purity of DNA extraction with sonication and spin column methods from the sputum sample of tuberculosis patient

Fitrianingsih Saputra; Asep Iin Nur Indra; Ai Djuminar; Fusvita Merdekawati; Betty Nurhayati

doi:10.29244/currbiomed.2.2.84-92

Fitrianingsih Saputra Jurusan Teknologi Laboratorium Medik, Poltekkes Kemenkes Bandung, Indonesia
Asep Iin Nur Indra Bidang Biologi Molekuler, Poltekkes Kemenkes, Bandung, Indonesia
Ai Djuminar Bidang Biologi Molekuler, Poltekkes Kemenkes, Bandung, Indonesia
Fusvita Merdekawati Bidang Biologi Molekuler, Poltekkes Kemenkes, Bandung, Indonesia
Betty Nurhayati Bidang Biologi Molekuler, Poltekkes Kemenkes, Bandung, Indonesia

DOI: https://doi.org/10.29244/currbiomed.2.2.84-92

Keywords: DNA concentration, DNA purity, tuberculosis sputum, sonication, spin column

Abstract

Background: The Polymerase Chain Reaction (PCR) method can identify Mycobacterium tuberculosis in a sputum sample of a patient with TB (TB). One crucial step to ensure accurate PCR results is the DNA extraction process. Objective: The research aims to compare the concentration and purity of DNA from the sputum of TB patients using ultrasound and spin column extraction techniques. Methods: The research uses descriptive study designs with post-only design strategies. The primary data was derived from 18 sputum specimens from TB patients. Concentration measurement and DNA purity testing using a nanodrop spectroscopic photometer. Results: DNA extraction by ultrasound method has an average concentration of 18.9 ± 8.5 ng/L, with a peak of 37.6 ng/ L. The spin column method produces an average of 55.5 ± 27.9 ng/μL; the peak is 105.0 ng/ μL. The purity value of the DNA extract is in the range of 1.8 ± 2.0 with the ultrasound method of 61% and the spin column of 78%. Conclusion: The sonication method has a lower average concentration and a higher percentage of purity than the spin column method, and there are differences in concentrations and purity values between the two methods.