Abstract
The isolation of DNA from biological samples is a crucial step in the process of DNA-based molecular biological assays. In this study, we present a simple method to extract and purify genomic DNA from different sample types (standard cyanobacteria, cyanobacteria in water samples and leaves of bamboo plants) which would be suitable for any molecular analyses. In this method, cyanobacteria are lysed by three sequential freezing and heating followed by enzymatic treatment with lysozyme and proteinase K. The extraction and purification of DNA was achieved with the lysing and nuclease inactivating properties of the chaotropic agent guanidium isothiocyanate and the nucleic acid binding properties of silica particles. The DNA extraction method yielded high quality reproducible DNA. The technique described here is a rapid, robust and cost effective method suitable for the extraction of DNA from any source for routine molecular biological assays. Key Words: Boom’s method, guanidine thiocyanate, silica, Polymerase Chain Reaction (PCR) DOI: http://dx.doi.org/10.4038/cjsbs.v40i1.3407 CJSBS 2011; 40(1): 59-63
Highlights
The isolation of DNA from biological samples is a crucial step in the process of DNAbased molecular biological assays
The technique described here is a rapid, robust and cost effective method suitable for the extraction of DNA from any source for routine molecular biology assay. This DNA extraction method yielded high quality DNA from different types of samples which were suitable for molecular analyses
DNA samples submitted to Polymerase Chain Reaction (PCR) reactions from the M. aeruginosa (PCC 7941), Lyngbya (PCC 8937) and the water samples collected from the environment for the 16S rRNA gene, yielded the fragment of about 450 bp, using the cyanobacterial specific oligonucleotide primers of Cya359F forward, Cya781 Ra and Cya781 Rb reverse (Fig. 1)
Summary
The isolation of DNA from biological samples is a crucial step in the process of DNAbased molecular biological assays. Whether the DNA is extracted from a plant or animal tissue or from a bacterium, the product obtained has to be pure or free from contaminants (proteins, carbohydrates) to be used in numerous applications in molecular biology including PCR, genotyping, DNA sequencing, etc. A wide variety of protocols are found in the literature to extract and purify genomic DNA from different tissues. All protocols start with cell lysis as the first step, followed by deproteination and precipitation of DNA. There are many different and versatile commercial kits suitable for genomic DNA extractions from QIAamp, Puregene and Dynabeads (Cler et al, 2006)
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