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The two constructs were stably transformed to Arabidopsis (Col-0) plants, and 20 independent transformants were collected and analyzed for each construct.
Two independent transformants were collected and displayed similar phenotypes (strains ΔBAR1-21 and ΔBAR1-31).
Cells were harvested and resuspended in YPD with or without salt (500 mM NaCl) for 30 min. Cells from three independent transformants were collected and assayed for β-galactosidase activity as described above.
Ampicillin-resistant transformants were collected and stored in 384-well plates.
A total of 6 × 10 yeast transformants were collected and pooled as the prey library.
Ampicillin-resistant transformants were collected and then stored in 384-format plates at −80°C.
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Transformant cells were collected from media by centrifugation and resuspended densely in PBS and deposited onto silane-coated slides (Sigma).
The crude extracts of WT and transformant cells were collected and dissolved in lysis buffer (1 ml of 40 mM Tris HCl pH 8.0) with protease inhibitor (1 mM phenylmethanesulfonyl fluoride).
To minimize positional effect of reporter gene expression, at least 1000 transformants on the membrane were collected to create a mixed population of transformants, which were cultured and assayed for the strength of GFP florescence.
Rooted primary (T0) transformants were transferred into soil, flowering plants were self-pollinated and T1 seeds were collected.
Seeds were collected for 5 individual transgenic lines and the screening of positive transformants was conducted as in [ 35].
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