Plant DNA extraction by CTAB Method

Описание: CTAB plant DNA extraction method
The CTAB DNA extraction method is simple and effective. Besides, the CTAB buffer, other ingredients are RNase, proteinase K, SDS.

Take 5gms of fresh plant tissue and cut it in the small pieces.
Add liquid nitrogen to the tissue and roughly grind the sample into the mortal and pastel.
After the tissue becomes a powder, add 500 μL of CTAB extraction buffer and beta-mercaptoethanol, grind it again followed by the vortexing for 5 minutes.
After it becomes homogeneous, take the mixture into the 2ml Eppendorf tube.
Boil the sample for 30minutes at 60°C to 65°C.
Centrifuge the sample at 10,000rpm for 5 to 8 minutes and transfer the supernatant into another tube.
Be careful, take the only supernatant, don’t mess the supernatant with the debris.
Now add 5 μL to 10 μL of RNase solution to the supernatant and incubate at 37°C for 25 minutes.
The proteinase K step is additional (you can use it if necessary).
Centrifuge the sample at 10,000rpm for 2 minutes and take supernatant to another tube.
Now add chilled isopropanol (70%), a pinch of NaCl to the supernatant and precipitate the DNA by inverting the tube several times.
Centrifuge the precipitate for 10,000rpm for 2 minutes and collect the pellet (remove the supernatant).
Now wash the DNA pellet with alcohol two times at 9000 rpm for 2 minutes.
After the clear pellet appears, add 500μL of TE buffer or elution buffer to the pellet and dissolve the DNA in it.
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