Three replicates of about 0.1 g embryo or 0.75 g endosperm
powder each were homogenized in 1.5 ml precooled extraction
buffer which contained 50 mM TriseHCl (pH 7.5), 30% (w/v) sucrose,
10 mM ethylene glycol-bis (b-aminoethylether)-N,N,N0,N0-
tetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1 mM
dithiothreitol (DTT) and 1% (v/v) Triton X-100. After washing the
mortar with 0.5 ml extraction buffer, the total of 2 ml homogenate
was centrifuged twice (at 16 000g at 4 C for 10 min and at 32 000g
Three replicates of about 0.1 g embryo or 0.75 g endospermpowder each were homogenized in 1.5 ml precooled extractionbuffer which contained 50 mM TriseHCl (pH 7.5), 30% (w/v) sucrose,10 mM ethylene glycol-bis (b-aminoethylether)-N,N,N0,N0-tetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1 mMdithiothreitol (DTT) and 1% (v/v) Triton X-100. After washing themortar with 0.5 ml extraction buffer, the total of 2 ml homogenatewas centrifuged twice (at 16 000g at 4 C for 10 min and at 32 000g
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