1. Picked Colonies of Existing BioBrick Transformants
3 colonies of E7, 2 colonies of E0, 2 colonies of R0, and numbered accordingly
2. Ran Gel of DNA Nanostructures
- Ran 2% agarose gel of the nanostructure reaction and the 2 controls for ~40 min at 130V.
1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold (NB: Only left four lanes are ours.)
1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA
- Three reactions were mixed according to given protocol.
oligos + scaffold -oligos -scaffold
2. Transformed Bacteria with Existing BioBrick Plasmids
- Transformed into OneShot Top10 competent cells (Invitrogen) according to the following protocol.
R0010 (lac operon promoter), E7104 (T4 promoter + gfp), and E0241 (gfp).
Thaw cells; aliquot out ~30uL per transformation. Add desired amount of plasmid DNA (1 uL in this case?) to cells and leave on ice for 30 min. Heat shock at 37°C for 30 sec in a heating block. Ice for 2 min. Shake in 37%deg incubator for 1 hr. Pour onto plates with the required antibiotic (carbinomycin in this case); spread with beads or plate spreader. Incubate plates in 37%deg incubator overnight.
HOMEWORK: 1) Past iGEM Projects Presentation: UCSF 2) Prospective Projects Research: a) Lactobacillus Hijacking b) Receptor-based DNA Nanostructure Latch c) Biocryptography w/ DNA Nanostructures d) ?