500x stock solution of nitrofurantoin (50 ml/ml) should be prepared by dissolving in dimethylformamide (DMF).Incubate the LB plate at 37 °C (optimal temperature for bacterial growth) overnight (~ 18 h).Day 2Streak the overnight-grown bacterial cells from a single colony on a CPG agar plate, using a metal inoculation needle . #####%%%%% output "bacterial growth" are used in line16
{"action": "prepare", "output": "bacterial growth", "reagent": ["500x stock solution of nitrofurantoin"], "volume": ["50 mg/ml"]}
Incubate the inoculated CPG agar plate at 30 °C (optimal temperature for the production of pigment, which is a main determinant for bacterial tolerance to UV) for 48 h (Figure 1).imgsrc:https://en-cdn.bio-protocol.org/attached/image/20160229/20160229061111_9425.pngFigure 1. #####%%%%% output "Burkholderia glumae" are used in line6
{"action": "incubate", "output": "Burkholderia glumae", "temperature": ["30 \u00b0"], "container": ["LB plate"]}
Strains of Burkholderia glumae grown on CPG agar plates.
{"action": "grow", "output": "", "reagent": ["Burkholderia glumae"]}
48 h of incubation at 30 °C.Day 4Resuspend the bacterial colonies grown on CPG agar in sterile water in a sterile microcentrifuge tube by flicking with hands.
{"action": "resuspend", "output": "", "reagent": ["bacterial colonies"], "device": ["sterile microcentrifuge tube"]}
Adjust the bacterial concentration to OD600 = 0.1 (~0.5 x 107 CFU/ml), using a spectrophotometer. #####%%%%% output "the diluted B. glumae cells" are used in line12
{"action": "adjust", "output": "the diluted B. glumae cells", "device": ["spectrophotometer"]}
Spread 100 µl of the diluted B. glumae cells on a CPG plate.
{"action": "spread", "output": "", "reagent": ["the diluted B. glumae cells"], "container": ["a CPG plate"]}
An image showing the UV irradiation procedure To determine the number of CFU accuratelyIncubate the inoculated plates at 30 °C for 48 h.Day 6Count the surviving bacterial colonies , serial dilutions up to 1:1,000 should be made (Figure 3).imgsrc:https://en-cdn.bio-protocol.org/attached/image/20160229/20160229062125_1328.pngFigure 3. #####%%%%% output "the surviving bacterial colonies" are not used
{"action": "count", "output": "the surviving bacterial colonies", "concentration": ["CFU"]}
spread on CPG agar plates
{"action": "spread", "output": "", "container": ["CPG agar plates"], "reagent": ["bacterial growth"]}
From left to right: Each vertical row represents the CPG agar plates exposed to UV for 0, 10, 20, , .
{"action": "provide", "output": "", "time": [""]}
Convert the data to percentage of survival in different time of exposure to UV radiation.
{"action": "convert", "output": ""}