hi,

(1)    Grind sample in LN (N2).

(2)    Add 500 µl Extraction Buffer (EB) + 10 µl mercaptoethanol.

(3)    Incubate at 65° C for 2-4 hours in water bath.

(4)    Cool and add 500 µl Chlorophorm: Isoamyl (24:1 vol:vol); and then spin it at 12000 rpm for 10-20 min.

(5)    Precipitate vol. of ice-cold Isopropanol (+ ⅟10 NaCl) to the aqueous phase.

(6)    Incubate at -20° C for 1-2 hours and then spin at 12000 rpm for 10 min.

(7)    Add wash buffer (100-200 ethanol + 3M NaCl).

(8)    Re-suspend in 100-200 high-salt TE buffer.

(9)    Add 3-10 RNase and incubate at 37 C for 30-40 min.

(10)Add Isopropanol and spin after cooling.

(11)Wash the pellet DNA with Ethanol.

(12)Dry and re-suspend.

(13)Repeat washing 2-3 times.

(14)Dry and re-suspend in 100 TE buffer.

-          Prepare 100 mL of Agaraose Gel.

Load the well (2 µl 6X loading buffer + 2 µl DNA sample).