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

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

DNA fragments werereceivedfrom Integrated DNA Technologies (IDT)

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

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

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

PCR primers werereceivedfrom BIOFACT™

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