Isolation_Of_Genomic_DNA_From_Marine_Fish_Using_Protein_Prec

Isolation Of Genomic DNA From Marine Fish Using Protein Precipitation Method. Introduction : DNA isolation is the very first step to study the genomics of a selected organism. It is crucial that the DNA extraction is carried out in a rapid, economical and simple manner. Various approaches are available for the scientists to isolate the genomic DNA e.g. cesium chloride density gradients, organic extraction method and salting out methods. In this lab work, the approach of salting out method is used and described, where different species of marine fishes are used as the sample. It involves a series of stages based on the manipulation of the solubility properties of the cellular components. This approach involves selective precipitation of the proteins rather than degradation of the proteins, which is less expensive, not using toxic chemicals; at the same time, is less specific, less reproducible and the DNA yield is highly variable. The basic principles shall be described in the discussion section. It is suggested that different combination of the organic extraction method and salting-out method may lead to higher concentration and better DNA yield. Materials and Reagents : 1. Extraction buffer - 100mM Tris pH8.0, 500mM NaCl 2%(w/v) SDS, which is freshly prepared. 2. 5M potassium acetate, pH6.5 3. 3M sodium acetate, pH7.6 4. 1005 isopropanol, -20oC 5. TE buffer: 10mM Tris pH8.0, 1mM EDTA pH8.0 6. 80% ethanol, -20oC 7. Micropestle 8. Aspirator dna, minutes, gel, tissue, buffer, v2, agarose, added, 4oc, volume, tube, solution, incubated, extraction, until, tris, supernatant, prepared, microfuge, m2v2, m1v1, concentration, centrifuged, casting, acetate, vacuum, transferred, sp, sds, sample, ph80, pellet, new, nacl, mixed