Research Article
Volume 18 Issue 7 - 2022
Modified Methods for Quick and Safe Extraction of DNA from Microbiological Samples
OO Ajayi1,2*, A Adekanmbi1, M Dianda2,3 and OE Fagade1
1Environmental Microbiology and Biotechnology Laboratory, University of Ibadan, Nigeria
2Soil Microbiology Unit, International Institute of Tropical Agriculture (IITA), Ibadan
3Laboratoire de Microbiologie Forestière (INERA/DEF)BP Ouagadougou, Burkina Faso
*Corresponding Author: OO Ajayi, Environmental Microbiology and Biotechnology Laboratory, University of Ibadan, Nigeria.
Received: May 06, 2022; Published: June 20, 2022




Abstract

Isolating and purifying DNA are very important steps in DNA molecular techniques used in microbiological studies for the identification of genotypes, traits associated with genes of interest, and genetic diversity. Plant and soil materials are among the most difficult for high-quality DNA extractions. While carrying out DNA extraction for a total of 2000 different sample types (including water, soil, plant, algae, fungi and bacteria) using two different kits (ZYMO extraction kit and E.Z.N.A.® soil DNA Kit) and the crude extraction cetyltrimethylammonium bromide (CTAB) method, several modifications were made to ensure extraction of good quality DNA. These modifications in the methods were documented, repeated to confirm their accuracy and were done without the use of liquid Nitrogen. The extracted DNA concentrations obtained from CTAB extraction were extremely high and were diluted for use in PCR using a ratio of 1:10 of DNA to water while those obtained from the kit were used directly (but in small quantities ~ 0.5μL) or diluted in a ratio of 50:50. The mean DNA yields obtained for CTAB extraction was between 500 - 4000 ng/μl while that obtained for the kits were between 70 - 400 ng/μl. The 260/ 280 nm absorbance ratio had a high level of purity between 1.8 and 2.0. These modified methods can be used for day to day extraction processes in the laboratory.

Keywords: Dna; Recovery; Extraction; Microbial; Efficient

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Citation: OO Ajayi., et al. “Modified Methods for Quick and Safe Extraction of DNA from Microbiological Samples”. EC Microbiology 18.7 (2022): 13-20.

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