Custer_2_3
To extract DNA from Cannabis floral organs, tissue (~1 g) was ground to fine powder, CTAB buffer (2.5ml, 1.5x) was added, then the mixture was incubated (65 for 30 min). Then, an equal volume of chloroform/isoamyl alcohol (24:1) was added, the solution was mixed well for 20 minutes, then centrifuged (3000 rpm for 20 min). The supernatant was transferred into a new tube and 0.1 volumes of CTAB (preheated to 65) and an equal volume of chloroform/isoamyl alcohol (24:1) was added, the solution was mixed well for 20 minutes, then centrifuged (3000 rpm for 20 min). The supernatant was transferred to a new tube and 2.5 volumes of 1% CTAB were added to precipitate the DNA. After 10 minutes, the DNA was centrifuged at 3000 rpm for 20 min, the supernatant was discarded, then NaCl (1ml, 1M) and 1 l RNase were added to dissolve the DNA at 65 overnight. The DNA was then precipitated by adding two volumes of isopropanol and incubating (-20 for 1h). The supernatant was removed, then the DNA was washed once with 75% ethanol, air dried, and finally dissolved in distilled water.