This Readership is 10 times more when compared to other Subscription Journals (Source: Google Analytics)
All submissions of the EM system will be redirected to Online Manuscript Submission System. Authors are requested to submit articles directly to Online Manuscript Submission System of respective journal.
Hydrocarbons are problematic environmental pollutants due to bioaccumulations in both soil and water environments and
slow biodegradation. Removal of these hydrocarbons is considered one of the main concerns with regards to bioremediation
activities. Identification of microbial hydrocarbon genes is vital to bioremediation strategies. Efficient extraction of high
molecular weight genomic DNA for downstream applications like next generation sequencing (NGS) and metagenomic library
construction presents a major challenge. A common problem with DNA obtained from soil is the presence of humic and fulvic
acids which co-extract with the DNA. Furthermore, traces of organic substances and heavy metals remain in the DNA extract
leading to interference with molecular methods downstream. In order to study the diversity of a complex environment like a
hydrocarbon-contaminated soil through culture independent techniques, efficient protocols that eliminate these contaminants
and yield good quality DNA must be developed. This study describes a modified method for extracting genomic DNA from heavy
oil contaminated soils. A general soil extraction method using CTAB was implemented however following the centrifugation of
the soil and CTAB extraction buffer after incubation at a temperature of 65 °C, the supernatant was filtered through two layers
of mesh guaze in order to remove the thick layer of oil floating at the top. Humic acid removal was also carried out using 2%
CaCl2 on the crude DNA. The isolated crude DNA was separated on an agarose gel and gel electroelution was carried out on the
excised gel fragments. Both the quality and quantity of isolated DNA was compared with existing methodologies and was found
to be of high quality and an acceptable quantity. The maximum recovery of genomic DNA in the absence of substantial amount
of impurities can be attributed to modifications in the isolation protocol and use of the gel electroelution technique.
Cindy Baburam is a Lecturer and Researcher in the Biotechnology Department at the Vaal University of Technology and is currently pursuing her PhD, working under the supervision of N A Feto. Her Master’s degree was conducted at the South African Sugarcane Research Institute in Durban, South Africa.