Methods for Plant Molecular Biology by Arthur Weissbach and Herbert Weissbach (Eds.)

By Arthur Weissbach and Herbert Weissbach (Eds.)

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5%. 0% (w/w) for each polymer, and the NaCl concentrations will range from 0 to 40 mM. Four stock solutions plus water and membranes are required (Table I). It is important to prepare the final experimental solutions by weighing them. 75 ml of 1 M sucrose in 20 mM 21 P. A. Albertsson, B. Anderson, C. Larsson, and H. E. Akerlund, Methods Biochem. AnaL2S9 115(1982). 22 C. Larsson, in "Moderne Methoden der Pflanzenanalyse" (H. F. Linskens and J. F. ), New Ser. Springer-Verlag, Berlin and New York, (in press).

5 A thorough comparison of the two procedures in the same study has not been made, thus it is difficult to assess which is superior. However, the sucrose gradient procedure appears to purify plasma membrane vesicles that are a mixture of right-sideout and inside-out vesicles that are mostly permeable to ions and MgATP910 whereas, the two-phase system appears to provide primarily a purification of right-side-out plasma membrane vesicles that are impermeable to Mg-ATP11. Since the purification procedure of choice may depend upon the planned use of the membranes, both procedures will be described.

These products are analyzed by GC and by gas chromatographymass spectrometry (GC-MS). 37 [1]CELL WALLS Identification and Quantitation of Partially O-Acetylated, Partially OMethylated Alditols. The positions of O-acetyl and O-methyl groups on the partially O-acetylated, partially O-methylated alditols are determined by GC-MS. GC-MS analysis is performed with a fused-silica, 30-m, SP 2330 column in the splitless mode. The following temperature program is used. Two minutes at an initial temperature of 80°, increased to 170° at 30°/min, then to 240° at 47min, and held for 5 min at 240°.

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