Table 1. Composition of chloroplast isolation buffers and wash buffers for modified high salt method, high salt plus saline Percoll method and sucrose gradient method.

High salt plus saline Percoll method	Modified high salt method [32]	Sucrose gradient [33]
Isolation Buffer (pH 3.8)	Isolation Buffer (pH 3.8)	Isolation Buffer
1.25 M NaCl	1.25 M NaCl	50 mM Tris-HCl (pH 8.0)
0.25 M ascorbic acid	0.25 M ascorbic acid	0.35 M sorbitol
10 mM sodium metabisulfite	10 mM sodium metabisulfite	7 mM EDTA
0.0125 M Borax	0.0125 M Borax	0.1% 2-mercaptoethanol
50 mM Tris-HCl (pH 8.0)	50 mM Tris-HCl (pH 8.0)	0.1% BSA
7 mM EDTA	7 mM EDTA
1% PVP-40 (w/v)	1% PVP-40 (w/v)
0.1% BSA (w/v)	0.1% BSA (w/v)
	1 mM DTT
Wash Buffer (pH 8.0)	Wash Buffer (pH 8.0)	Wash Buffer
1.25 M NaCl	1.25 M NaCl	50 mM Tris-HCl (pH 8.0)
0.0125 M Borax	0.0125 M Borax	0.35 M sorbitol
50 mM Tris-HCl (pH 8.0)	50 mM Tris-HCl (pH 8.0)	25 mM EDTA
25 mM EDTA	25 mM EDTA
1% PVP-40 (w/v)	1% PVP-40 (w/v)
0.1% BSA (w/v)	0.1% BSA (w/v)
	1 mM DTT

Both BSA and DTT were added just before the start of the experiment.

Percoll gradient solutions consisted of wash buffer with Percoll at a final concentration of 70% (v/v) and 30% (v/v).

Sucrose gradient solutions consisted of 50 mM Tris-HCl (pH 8.0), 25 mM EDTA and sucrose addition for a final concentration of 52% sucrose (w/v) and 30% (w/v) sucrose.