PCR and Western blots confirmed that the transformant was fully segregated and all HoxF in the mutant was linked to GFP (ESI Fig. 2 and 3 †).
They were fully prepped".
A total of 40 independent primary transformants were selected and detached leaves were inoculated with P. infestans isolate 90128; leaves from 34 plants remained resistant, whereas those from wildtype Desiree control plants were fully susceptible.
All transformants were homozygous for a single insertion and have been fully characterized in previous publications.
Southern gel blot analysis confirmed that all T3 lines were single-copy independent transformants and all have been fully characterized [40], [53].
This fragment was transformed into WT Synechocystis and transformants were selected on BG-11 agar plates containing 2 μg/ml zeocin and fully segregated by incrementally doubling the concentration of antibiotic to 16 μg/ml.
Furthermore, transformation protocol for generating a large number of transformants is limited to only a few cultivars, indicating that experimental parameters for rice transformation have not been fully optimized (Ozawa 2009).
Transformants were selected on kanamycin-containing LB agar plates.
The transformants were selected on the basis of green fluorescence, total genomic DNA (gDNA) was extracted from these fluorescent transformants.
