User:TChan: Difference between revisions
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==W 6.14.06== | ==W 6.14.06== | ||
1. | 1. Miniprep of the 3 BioBricks Plasmids (according to [http://openwetware.org/wiki/Miniprep/Qiagen_kit standard protocol]) | ||
2. Double digest of the 2 R0 (lac promoter) and 2 E0 (gfp) samples | |||
3. Phosphatasing of 2 R0 samples to prevent self-ligation | |||
4. Ran R0 and E0 samples on gel to separate out the gfp from the E0-plasmid | |||
5. Gel extracted R0 plasmid (~2.1 kb, linearized by digestion) and E0 gfp fragment (~0.9kb) | |||
==Tu 6.13.06== | ==Tu 6.13.06== | ||
1. Picked Colonies of Existing BioBrick Transformants | 1. Picked Colonies of Existing BioBrick Transformants |
Revision as of 14:13, 14 June 2006
W 6.14.06
1. Miniprep of the 3 BioBricks Plasmids (according to standard protocol)
2. Double digest of the 2 R0 (lac promoter) and 2 E0 (gfp) samples
3. Phosphatasing of 2 R0 samples to prevent self-ligation
4. Ran R0 and E0 samples on gel to separate out the gfp from the E0-plasmid
5. Gel extracted R0 plasmid (~2.1 kb, linearized by digestion) and E0 gfp fragment (~0.9kb)
Tu 6.13.06
1. Picked Colonies of Existing BioBrick Transformants
- Picked:
3 colonies of E7, 2 colonies of E0, 2 colonies of R0, and numbered accordingly
- Grew each colony in 5mL LB + 5uL of 50mg/mL Amp (1:100 dilution).
2. Ran Gel of DNA Nanostructures
- Ran 2% agarose gel of the nanostructure reaction and the 2 controls for ~40 min at 130V. (NB: Only left four lanes are ours.)
1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold
M 6.12.06
1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA
- Three reactions below were mixed according to given protocol.
oligos + scaffold -oligos -scaffold
2. Transformed Bacteria with Existing BioBrick Plasmids
- Transformed each BioBrick plasmid below into OneShot Top10 competent cells (Invitrogen) according to the following protocol.
R0010 (lac operon promoter), E7104 (T4 promoter + gfp), and E0241 (gfp).
Top10 Transformation
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:
- Past iGEM Projects Presentation: UCSF
- Prospective Projects Research:
Lactobacillus Hijacking Receptor-based DNA Nanostructure Latch Biocryptography w/ DNA Nanostructures ?
Contact Info
/Tiffany Chan /Cell: 781 330 1969 /chan.tiffany at gmail.com