EFFICIENT AND PRACTICAL APPROACH FOR DNA EXTRACTION
Presenter: IZHAR ALI & BABAR USMAN
Ph.D. ScholarsGUANGXI UNIVERSITYNanning, P.R. China
Purpose of DNA Extraction
- To obtain DNA in a relatively purified form which can be used for further investigations, i.e. PCR, sequencing, etc.
Note: ( i will upload full video after sometime so you may learn from this blog) and you can download this file in world Click Here
Procedure & Protocol

- Take 1 gm of leaf sample.
- Pulverize plant tissue in liquid nitrogen using a mortar and pestle.
- Take the grinded material in Eppendorf tubes.
- Add 800 μl CTAB-Cetyl trimethyl ammonium bromide.
- Keep the tubes in water bath at 65 °C for 35 minutes and shake three times after each 10 minutes.
- After taking sample from water bath, allow the sample to cool at normal temperature.
- Add 800 μl of 24:1 CIA (Chloroform Isoamyl Alcohol).
- Gently shake the sample.
- Centrifuge the sample at 1300 rpm for 10 minutes.
- After centrifuge take 550 μl supernatant from the tubes and discard the debris.
- Again add 550 μl of 24:1 CIA.
- Shake the sample gently.
- Again centrifuge for 6 minutes at 12000 rpm.
- Take supernatant and discard the lower layer.
- Add chilled alcohol (Isopropanol) two times than your sample.
- Keep the sample in ice box/freezer for 30 minutes.
- Again centrifuge for 5 minutes.
- Discard the liquid material.
- Wash the pellet two times with 800 μl of 75% ethanol.
- Keep the tube on tissue paper for 30 minutes/over night.
- Add 50 μl of ddH2O.
- Keep the material in normal temperature for further use.
- Take 1 gm of leaf sample.
- Pulverize plant tissue in liquid nitrogen using a mortar and pestle.
- Take the grinded material in Eppendorf tubes.
- Add 800 μl CTAB-Cetyl trimethyl ammonium bromide.
- Keep the tubes in water bath at 65 °C for 35 minutes and shake three times after each 10 minutes.
- After taking sample from water bath, allow the sample to cool at normal temperature.
- Add 800 μl of 24:1 CIA (Chloroform Isoamyl Alcohol).
- Gently shake the sample.
- Centrifuge the sample at 1300 rpm for 10 minutes.
- After centrifuge take 550 μl supernatant from the tubes and discard the debris.
- Again add 550 μl of 24:1 CIA.
- Shake the sample gently.
- Again centrifuge for 6 minutes at 12000 rpm.
- Take supernatant and discard the lower layer.
- Add chilled alcohol (Isopropanol) two times than your sample.
- Keep the sample in ice box/freezer for 30 minutes.
- Again centrifuge for 5 minutes.
- Discard the liquid material.
- Wash the pellet two times with 800 μl of 75% ethanol.
- Keep the tube on tissue paper for 30 minutes/over night.
- Add 50 μl of ddH2O.
- Keep the material in normal temperature for further use.

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