Team:William and Mary/Notebook
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Week 1 (160529 – 160604)
✻ Resuspended parts from the kit.
✻ Resuspended gBlocks of ordered sequences.
✻ Created a functional UNS Standard Backbone, containing the UNS 2 and 3 sequences within the Prefix and Suffix.
✻ Cloned K2066001-K2066016 into UNS pSB1X3 Backbone.
✻ Performed Diagnostic PCRs to test primer design for Ribozyme / RiboJ Characterization subproject
✻ Transformed interlab devices, created glycerol stocks. Could not get IMP #1, #3, and (-) control to transform.
Week 2 (160605 – 160611)
✻ Constructed K2066024, K2066025, and K2066027 using Gibson Assembly (from K2066014, K2066015, and pTAC templates).
✻ We received training on the FACS (Fluorescence Activated Cell Sorter) Machine.
✻ Realized the need for repressor protein / fluorophore fusion proteins (LacI-mCherry, TetR-GFP, i.e.)
✻ Designed gBlocks of K2066028 and K2066029.
✻ Attempted to clone Addgene 240x TetO Sequence into UNS Standard Backbone.
✻ Attempted ICA with K2066002, K2066003, K2066004 to create a TetO w/ 8bp spacer 9-mer
✻ Constructed K2066030 and K2066031 from K2066015.
✻ Troubleshot ICA, attempted 6- and 12-mers of TetO w/ 8bp spacer.
✻ Attempted IPTG Induction of K2066014 + K2066016 cotransformations.
✻ Transformed remaining interlab devices, created glycerol stocks.
Notebook
