Extraction of DNA and RNA from Microorganism (original) (raw)

The enzyme lysozyme is used for the rupturing of cell wall of bacteria or microorganism. The high salt concentration prevents the conversion of dsDNA to ssDNA. EDTA and sodium citrate present helps to chelate metal ions which activate DNase and RNase enzymes. The protein denaturation is done by sodium chlorate and finally extraction is done either by ethanol or isopropanol method as discussed earlier in section. Material and Reagents: Any bacteria (for the first timers, use Escherichia coli). Solution A: Saline EDTA pH 8.0 in distilled water. Sodium chloride (0.15 mol/liter) ¼ 125 ml. EDTA (0.1 mol/liter) ¼125 ml. Enzyme lysozyme (N-acetylmuramide glycanhydrolase) ¼ 100 mg. Sodium lauryl sulfate (10% w/v) ¼ 50 ml. Sodium chlorate (6 mol/liter) ¼ 100 ml. Chloroform: Isoamyl alcohol (24: 1) ¼ 500 ml. Absolute ethanol. Sodium acetate (3 mol/liter) ¼ 25 ml. Isopropanol ¼ 100 ml. Sodium hydroxide (6 mol/liter) ¼ 50 ml.