Editing Agrobacterium tumefaciens transformation (section)

==  Arabidopsis growth and seed handling ==

See [http://www.biosci.ohio-state.edu/pcmb/Facilities/abrc/handling.htm#Growth_growth The Arabidopsis Biological Resource Center (ABRC)] for helpful hints on growing Arabidopsis.

'''Soil, fertilizer, pots, watering'''



#'''Soil:''' We use BX soil mix.
#'''Fertilizer:''' Prepare Miracle-Grow Liquid Houseplant Food (8-7-6) by mixing two capfuls of fertilizer per gallon of water.
#'''Insecticide:''' Marathon granular insecticide -- add 5g to the bottom of each flat upon initial sewing.
#'''Pots:''' To start seedlings use 3x5" tray inserts. For transplanting and large-scale growth use 36-pot inserts. Use 3" pots per plant for physiology/phenotyping experiments.

'''Starting seedlings'''



#Sprinkle granular insecticide on the bottom of the plant tray in which the pots will be placed.
#LOOSELY fill pots or flat inserts with BX soil mix. Lightly wet the top of the soil with tap water and fill bottom of tray with ~1/2 - 1" deep of fertilizer water. Once most of the water is absorbed, seeds can be sewn.
#To start seedlings, sprinkle ~50-100 seed per 3x5" pot and place in growth chamber or on light racks in lab. Seedlings are usually ready for transplant after 7-10 days.
#Water plants from the bottom only when the soil is starting to become dry, usually every 3-5 days. '''DO NOT OVERWATER OR LET PLANTS STAND IN WATER FOR EXTENDED PERIODS!''' Dump out excess water if it is not soaked up within an hour or so. Approximately 1/2" of water is appropriate for one flat.

'''Transplanting seedlings'''

#Prepare trays and soil as for starting seedlings.
#Once the soil is thoroughly wet, create a small hole ~1/2" deep with the end of a pen or sharpie. Gently transplant individual seedlings to individual pots and gently pack soil around roots.
#Fertilize flats during the initial transplanting and every two weeks thereafter with Miracle-Grow Liquid Houseplant Food (8-7-6) by mixing two capfuls of fertilizer per gallon of water. Fertilize as you would for standard watering (see below).
#Water plants from the bottom only when the soil is starting to become dry, usually every 3-5 days. '''DO NOT OVERWATER OR LET PLANTS STAND IN WATER FOR EXTENDED PERIODS!''' Dump out excess water if it is not soaked up within an hour or so. Approximately 1/2" of water is appropriate for one flat.