July
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7/9/2018
Measurement of Abs 600 and OD 600 for LUDOX CL-Xand water
Prepared a dilution series of silica microspheres and measure the Abs 600 in a plate reader
Prepared a dilution series of fluorescein in four replicates and measure the fluorescence
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7/10/2018 – 7/22/2018
Transform E. coli DH5α with InterLab study plasmids
Pick colonies from each of the transformation plates and inoculate in 5-10 mL LB medium +
Chloramphenicol. Grow the cells overnight. Cell growth, sampling, and assay
Repeat experiments were conducted to confirm the reproducibility of the data
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7/23/2018 (Composite parts cloning started)
DNA fragments werereceivedfrom Integrated DNA Technologies (IDT)
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7/24/2018 – 7/31/2018
Conducted A-tailing process at the end of our gene fragment for TA cloning
A-tailed gene fragment was ligated with the TA vector
TransformedE. coli DH5α with ligated TA vector (Blue-white screen)
Picked colonies from transformation plates. Cells were grown in LB broth with 0.1mM of ampicillin
Vector isolation was conducted through DNA mini prep kit
DNA fragment parts were purified using gel electrophoresis and ligated with pSB1C3 vector.
Checked appropriate insertion of the construct in the vector
Repeat experiments were conducted for debugging
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August
8/1/2018 – 8/11/2018
Plasmid sequence was confirmed by BIOFACT™ sequencing analysis service
Amplification and purification of GBP-ProG fusion Protein
Purified proteins were confirmed by SDS-PAGE
Protein quantified using the Bradford protein assay
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8/12/2018 – 8/19/2018
Plasmid sequence was confirmed by BIOFACT™ sequencing analysis service
Amplification and purification of GBP-ProG fusion Protein
Purified proteins were confirmed by SDS-PAGE
Protein quantified using the Bradford protein assay
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September
9/10/2018 (Basic parts cloning started)
PCR primers werereceivedfrom BIOFACT™
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9/11/2018 – 9/22/2018
A series of experiments was the same as a composite parts cloning experiment.
Conducted cloning experiments for two basic parts (GBP, ProG)
Plasmid sequence was confirmed by BIOFACT™ sequencing analysis service
Repeat experiments were conducted for debugging
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