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文献和实验Plant DNA methylation analysis by bisulfite genomic sequencing
(12000 X g); Discard supernatant, vacuum dry for 10 minutes at room temperature; Dissolve cut DNA in 10µl bidist water (0.25 or 2.5 µg/µl, respectively), store at -20°C; Bisulfite treatment The bisulfite solution
In Situ Hybridization Using Digoxigenin Labeled Probes
(I prefer overnight but if you're in a hurry, 3 hours should give a nice probe) 5) Add 20 ug carrier DNA and 1/10 volume 3M NaOAc. Precipitate with 2.5 volumes EtOH for 1 hr @ -70oC or overnight @ -20oC 6) Spin DNA in microfuge 15 min. Remove EtOH
wash 3x with 1ml EBC-buffer 9. resuspend beads in 100µl EBC-buffer and add 35µl 4x sample buffer (bound fraction) or to load total amount of bound fraction, add 30µl 1x sample buffer, boil 10min at 100°C put at -20°C
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