CTAB Extraction Protocol

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					CTAB Extraction Protocol 1. 2. 3. 4. Collect cells on GF-C filter Add 2 mL of 2X CTAB buffer (add 1 mL of CTAB to the CTAB/filter) Grind filter with wooden applicator sticks Incubate for 1 hour @ 60-65 °C, vortexing periodically (no more than 10 s at a time) 5. Add equal volume of 24:1 chloroform:isoamyl alcohol, mix thoroughly in 25 mL H2O. Add 2 mL to spec tube. 6. Centrifuge in clinical centrifuge for at least 20 min @ full speed or at 2700 rpm for 8 min. 7. Pipette top aqueous layer into 1.5 mL eppendorf tubes (approx. 850 μL x 2) 8. Add 0.7 volumes of 100% isopropanol (600 μL for 850 μL of soln) and invert tubes several times. 9. Centrifuge 25 mins @ max speed in a microfuge 10. Pour off supernatant and allow to air dry 11. Resuspend in 20 μL of sH20 or 25 μL TE.


				
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