Developing fruit flavedo at 165 DAFB

Organ: Fruit; Vector: pTriplEx2; Site_1: SfiIA; Site_2: SfiIB; Developing citrus fruits were harvested from trees growing in the Citrus variety collection in the Wolfskill experimental orchard located in Winters, California (USA). Fruit was collected on October 22, 2003, between 8 to 9 am and stored at 4C. The flavedo tissue was dissected out of developing fruit (165 DAFB) and used to isolate RNA using Trizol reagent from Invitrogen. The cDNA Library was constructed using the SMART cDNA library Kit (Clontech). The primary library was en masse evicted and plasmid DNA containing the cDNA library was isolated from the resultant bacterial population. Plasmid DNA was then transformed into ultra competent E coli cells (XL10 Gold; Stratagene). Transformants were plated out on Q-trays (2000 cfu/tray), picked using a Qbot and archived in 384 well dishes.