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Table 1 Overview of nuclei isolation protocols A, B and C

From: Isolation and analysis of high quality nuclear DNA with reduced organellar DNA for plant genome sequencing and resequencing

  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) Peterson et. al. [16] (Option X); Peterson et. al. [17]
  1. * For L. gibba and S. polyrhiza the volume of buffer A had to be doubled for isolation on non-degraded DNA.