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a-c Chloroplasts in the WT leaf (a), the yellow sector of an fgl leaf (b), and the OPAO #11 homozygous T2 line leaf (c) grown for 80 days in the greenhouse.
Per line, leaf material from on average 12 BC3S6 plants was pooled.
Each line was then thinned out to two 'healthy' plants per M2 line, leaf samples were taken for DNA isolation, and the plants were bagged individually and allowed to grow to maturity.
Briefly, 1 g of NRPB2-FLAG transgenic line leaf tissue or 4 g of NRPD1-FLAG transgenic line leaf tissue (for Pol IV) was ground to a powder in liquid nitrogen using a mortar and pestle and then resuspended in 3.5 ml extraction buffer (20 mM Tris-HCl, pH 7.6, 300 mM sodium sulfate, 5 mM magnesium sulfate, 5 mM DTT, 1 mM PMSF, 1% plant protease inhibitor cocktail [Sigma]) per gram of tissue.
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Everyone's favourite stoner, Snoop Dogg, launched his weed line Leafs by Snoop last week at a private party in Denver, and Willie Nelson is working on a line of weed and other THC products called Willie's Reserve.
After regaining possession at Michigan's 47-yard-line, Leaf threw a 15-yard touchdown pass to McKenzie to take a 7 0 lead with 3 07 remaining in the first quarter.
In the case of lines 231 and 308 lines leaf color of both diploids and tetraploids remained green (Fig. 2) indicating strong silencing of CHS by the transgene in these lines.
In order to obtain homotransplastomic lines, leaf explants from the regenerated plants were subcultured again on selective RMOP medium.
At the vegetative stage, the wild-type and CsHMA3 transgenic lines had 4.81 ± 0.16 and 3.68 ± 0.03 (mean ± SE of four transgenic lines) leaf index values, respectively.
To determine the relative expression of the different lines, leaf sections from fully expanded leaves were cut and stained for GUS activity.
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