DNA Extraction

 

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.