We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
GENOMIC DNA EXTRACTION METHODS: A COMPARATIVE CASE STUDY WITH GRAM-NEGATIVE ORGANISMS.
- Authors
JAHAN, Sanjida; CHOWDHURY, Sayeeda Fahmee; MITU, Shahida Akter; SHAHRIAR, Mohammad; BHUIYAN, Mohiuddin Ahmed
- Abstract
The cell wall structure of gram-negative bacteria is more complex than that of gram-positive bacteria because of the presence of two layers external to the cytoplasmic membrane. Purpose of this study is to compare the four different methods for genomic DNA extraction from three gram negative strains-Escherichia coli, Klebsiella species, and Salmonella typhi. The genomic DNA was extracted from gram-negative bacteria cultures using enzymatic lysis with Proteinase-K, chemical lysis with cetyl trimethyl ammonium bromide (CTAB), physical lysis with Triton X-100 and Beads beating methods. The yield and quality of DNA were compared by spectrophotometry and agarose gel electrophoresis. The genomic DNA purity ratio was found within 1.1-1.9 for all four methods. This study revealed that the chemical lysis with CTAB was the most efficient method for extracting genomic DNA from gram negative strains. In terms of chemical lysis with CTAB for Salmonella typhi and Klebsiella species the highest purity ratio were 1.9 and 1.8 respectively. However Escherichia coli also showed the highest purity ratio (1.7) for CTAB extraction. Enzymatic lysis with Proteinase-K was also very effective and purity ratio for Salmonella typhi and Klebsiella species were 1.8 and 1.7 respectively.
- Subjects
CETYLTRIMETHYLAMMONIUM bromide; GENOMIC imprinting
- Publication
Banat's Journal of Biotechnology, 2015, Vol 6, Issue 11, p61
- ISSN
2068-4673
- Publication type
Article
- DOI
10.7904/2068-4738-VI(11)-61