Team:UST Beijing/Project
Wiki of InterLab by UST_Beijing
Background
“All of the 2017 iGEM teams are invited and encouraged to participate in the Fourth International InterLaboratory Measurement Study in synthetic biology.” The goal of our team is to use the InterLab protocol and Measurement Kit provided by iGEM official to do measurements which can help to establish a GFP measurement protocol based on engineering principles that anyone with a plate reader can use in their lab. The specific task for us is to measure GFP fluorescence and standard curve by using plate reader.
Materials & Methods
Plasmids used
Kit Plate 7
Positive Control (BBa_I20270): well 21B
Negative Control (BBa_R0040): well 21D
Test Device 1 (BBa_J364000): well 21F
Test Device 2 (BBa_J364001): well 21H
Test Device 3 (BBa_J364002): well 21J
Test Device 4 (BBa_J364003): well 21L
Test Device 5 (BBa_J364004): well 21N
Test Device 6 (BBa_J364005): well 21P
Strain used
Escherichia coli DH5α
Materials
FITC Standard: one tube with dried down FITC for creating a FITC standard
• LUDOX: one tube with 30% colloidal silica suspended in 1mL of water
• 1xPBS (phosphate buffered saline)
• LB (Luria Bertani) media
• Chloramphenicol (stock concentration 25 mg/mL dissolved in EtOH)
• 50 ml Falcon tube (or equivalent) or 250 ml shake flask for cell growth
• 1.5 ml eppendorf tubes for sample storage
• Ice bucket with ice
• Pipettes
• 96 well plate
Machines
Enspire 2300-001M
Sigma Laborzentrifugen
Thermo Scientific Nanodrop 2000
EVOS AMG microscope
Software
Microsoft Excel
Methods
Calibration
OD600 Reference point
FITC fluorescence standard curve
Test Device 1 (BBa_J364000): well 21F
Test Device 2 (BBa_J364001): well 21H
Test Device 3 (BBa_J364002): well 21J
Test Device 4 (BBa_J364003): well 21L
Test Device 5 (BBa_J364004): well 21N
Test Device 6 (BBa_J364005): well 21P
Cell measurement
Transformation
Measurements
Protocols
Please click the link:
https://2017.igem.org/Competition/InterLab_Study>
