Hi,
Can anyone tell if there is any published method for isolation of genomic DNA from cotton lint fibres.
If not, can anyone give an insight for any work being done anywhere in the world on cotton lint DNA?
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).
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