Table 1 Overview of nuclei isolation protocols A, B and C
| Â | Protocol A | Protocol B | Protocol C |
|---|---|---|---|
Plant criteria | •Low amount of plant material available •Low levels of secondary metabolites | •DNase-rich plants •Large amounts of plant tissue available | •High levels of secondary metabolites |
Successful DNA isolation | •A. thaliana •B. distachyon •G. aurea •S. bicolor •L. gibba* •S. polyrhiza* •Z. mays | •B. distachyon •L. gibba •S. bicolor •S. polyrhiza •Z. mays | •V. macrocarpon |
Protocol Modifications | •Used only 0.25 g tissue •Only performed DNA isolation steps (6-13) •Removed protease inhibitors from all buffers •Resuspended nuclei pellet in TE (10 mM:1 mM) | •Omitted TE slurry and diethyl ether steps •Increased volume of SEB to 200 ml •Added Triton X-100 drop by drop to minimize disruption of the nuclear membrane •Isolated DNA by isopropanol precipitation | •Omitted TE slurry and diethyl ether steps •Added EGTA and L-Lysine-HCl to MEB Buffer •Incubated sample on ice 8 minutes after addition of Triton X-100 •Centrifuged filtrate one time •Resuspended nuclei pellet in 1 ml of MPDB per gram of starting tissue |
Protocol modified from | Gendrel et. al. [15] | Peterson et. al. [16] (Option Y) |