1. Grind young leaf tissues. Place about 50ul ground tissues into 2ml microtubes. Add 750 ul of 2x CTAB buffer and 50 ul of 20% SDS. Mix thoroughly.
                 Incubate at 65 oC for 30 minutes to 1 hour. Agitate every 15 minutes to homogenize buffer with ground tissues.

            2. Cool briefly and add 800 ul chloroform-isoamyl alcohol (24:1). Mix thoroughly. Spin at 10,000 rpm for 15 minutes.

            3. Carefully transfer the aqueous phase (top phase) into a new 1.5 ml tubes.

            4. Add 800 ul isopropanol, mix thoroughly and incubate at -20oC for 30 minutes or overnight.

            5. Spin at 10,000 rpm for 10 minutes. Decant isopropanol and wash pellet with 500 ul 70% ethanol, spin for 5 minutes, and then drain dry.

            6. Dissolve pellet in 100 ul TE + RNAse mix (100 ul TE: 1ul RNASE 10mg/ml) and incubate at 37oC for 30 minutes.


            7. Add 10ul 3M NaoAc and 200ul absolute ethanol. Mix and precipitate for 3 hours or overnight.

            8. Centrifuge at 10,000 rpm for 10 minutes and decant supernatant.

            9. Add 200 µl of 70% Ethanol and spin for 10,000 rpm for 5 minutes. Decant supernatant and dry pellets.

            10. Add 50 µl TE and mix until the pellet is dissolved.


            1. Prepare 1% agarose (e.g. 2.5g agarose in 250ml 0.5x TBE).

            2. Dispense 2µl of homogenized DNA samples and 4µl of 2x BJ in each well of a re-used plate. Spin down.

            3. Dispense samples in the wells of the gel. Add 2µl of 50ng/µl Lambda at the end of the gel.

            4. Run the gel in 150V for 30 minutes.


  •     2X CTAB (1liter)
                20g CTAB dissolve in 860ml sterile ddH2O
                    Add: 81.82g NaCl
                               100 ml 1M Tris pH8.0
                               40 ml 0.5M EDTA pH 8.1
                               Autoclave and store at room temp

  • 20% Sodium Dodecyl Sulfate (SDS)
                    Dissolve 20 g SDS in 100ml of sterile nanopure H2O.
                    Do not autoclave.

  • 3M Sodium Acetate
                    Dissolve 123.1g NaOAc in 400ml of dsH2O.
                    Adjust the pH to 6.0 and volume up to 500ml

  • TE Buffer (250 ml)
                    1M Tris (pH 8.0) 2.5 ml
                    0.5 M EDTA (pH 8.0) 0.5 ml
                    Sterile nanopure water 247.0 ml