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| - | ==W 6.14.06== | + | <div class="tabs-blue"> |
| | + | <ul> |
| | + | <li id="current">[[TChan|Project Overview]]</li> |
| | + | <li>[[TChan/Schematics|Schematics]]</li> |
| | + | <li>[[TChan/OligoList|Oligo List]]</li> |
| | + | <li>[[TChan/Scaffold|Scaffold]]</li> |
| | + | <li>[[TChan/Images|Images]]</li> |
| | + | <li>[[TChan/iGEMNotes|iGEM Notes]]</li> |
| | + | </ul> |
| | + | </div> |
| | + | <br style="clear:both"> |
| | + | <br> |
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| - | <b>Goal:</b> Digest and extract out the E0-gfp and ligate after the R0-promoter, into that pre-cut plasmid.
| + | *See [[IGEM:Harvard/2006/DNA_nanostructures|iGEM 2006 Page]] for more info |
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| + | |
| - | R0: ---|E--|X---lacprom---|<b>S</b>--|<b>P</b>---
| + | |
| - | E0: ---|E--|<b>X</b>-----gfp-----|S--|<b>P</b>---
| + | |
| - | ---
| + | |
| - | hypothetical ligation:
| + | |
| - | ------|E--|X---lacprom---|M-----gfp-----|P---
| + | |
| | | | |
| - | 1. Miniprepped the 3 BioBricks Plasmids (according to [http://openwetware.org/wiki/Miniprep/Qiagen_kit standard protocol])
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| - | *Made glycerol stocks of 1mL of the cultures from each transformant:
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| - | 666.6uL of 50% glycerol (to give 20% glycerol total)
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| - | 1mL of transformant culture
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| - | 2. Double digested the 2 R0 (lac promoter) and 2 E0 (gfp) samples
| + | ==iGEM 2006 Notebook== |
| - | *NB: Due to the NanoSpec reading out negative DNA concentrations, we proceeded with an arbitrary amount of transformed DNA for the digests.
| + | <calendar> |
| - | *Digested with SpeI and PstI (NEBuffer 2) for R0, XbaI and PstI (NEBuffer 3) for E0, with the following protocol:
| + | name=IGEM:Harvard/2006/DNA_nanostructures/Notebook |
| | + | date=2006/07/01 |
| | + | view=threemonths |
| | + | format=%name/%year-%month-%day |
| | + | weekstart=7 |
| | + | </calendar> |
| | | | |
| - | ===DIGEST PROTOCOL=== | + | <calendar> |
| - | *For a total volume of 25 uL:
| + | name=IGEM:Harvard/2006/DNA_nanostructures/Notebook |
| | + | date=2006/10/01 |
| | + | view=threemonths |
| | + | format=%name/%year-%month-%day |
| | + | weekstart=7 |
| | + | </calendar> |
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| - | 8
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| - | 3. Phosphatasing of 2 R0 samples to prevent self-ligation
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| - | 4. Ran R0 and E0 samples on gel to separate out the gfp from the E0-plasmid
| + | ==Notebook== |
| | + | <calendar> |
| | + | name=TChan/Notebook |
| | + | date=2006/2/01 |
| | + | view=threemonths |
| | + | format=%name/%year-%month-%day |
| | + | weekstart=7 |
| | + | </calendar> |
| | | | |
| - | 5. Gel extracted R0 plasmid (~2.1 kb, linearized by digestion) and E0 gfp fragment (~0.9kb) | + | <calendar> |
| | + | name=TChan/Notebook |
| | + | date=2006/5/01 |
| | + | view=threemonths |
| | + | format=%name/%year-%month-%day |
| | + | weekstart=7 |
| | + | </calendar> |
| | | | |
| - | ==Tu 6.13.06==
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| - | 1. Picked Colonies of Existing BioBrick Transformants
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| - | *Picked and numbered:
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| - | 3 colonies of E7,
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| - | 2 colonies of E0,
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| - | 2 colonies of R0.
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| - | *Grew each colony in 5mL LB + 5uL of 50mg/mL Amp (1:100 dilution).
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| - | 2. Ran Gel of DNA Nanostructures
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| - | *Ran 2% agarose gel of the nanostructure reaction and the 2 controls for ~40 min at 130V. (NB: Only left four lanes are ours.)
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| - | [[Image:Tc_mm_pt_lh_folding_0613.jpg|thumb|left|Nanostructure Gel]]
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| - | 1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold
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| - | ==M 6.12.06==
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| - | 1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA
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| - | *Three reactions below were mixed according to [http://openwetware.org/wiki/Folding_DNA_nanostructures given protocol].
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| - | oligos + scaffold
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| - | -oligos
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| - | -scaffold
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| - |
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| - | 2. Transformed Bacteria with Existing BioBrick Plasmids
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| - | *Transformed each BioBrick plasmid below into OneShot Top10 competent cells (Invitrogen) according to the following protocol.
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| - | R0010 (lac operon promoter),
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| - | E7104 (T4 promoter + gfp),
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| - | and E0241 (gfp).
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| - |
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| - | ===Top10 Transformation===
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| - | Thaw cells; aliquot out ~30uL per transformation.
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| - | Add desired amount of plasmid DNA (1 uL in this case?) to cells and leave on ice for 30 min.
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| - | Heat shock at 37°C for 30 sec in a heating block.
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| - | Ice for 2 min.
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| - | Shake in 37%deg incubator for 1 hr.
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| - | Pour onto plates with the required antibiotic (carbinomycin in this case); spread with beads or plate spreader.
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| - | Incubate plates in 37%deg incubator overnight.
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| - | HOMEWORK:
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| - | # Past iGEM Projects Presentation: UCSF
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| - | # Prospective Projects Research:
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| - | Lactobacillus Hijacking
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| - | Receptor-based DNA Nanostructure Latch
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| - | Biocryptography w/ DNA Nanostructures
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| - | ?
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| | ==Contact Info== | | ==Contact Info== |
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| | Biochemical Sciences | | Biochemical Sciences |
| | Kirkland '07 | | Kirkland '07 |
| - | PRISE Summer 2006 | + | Academic Interests: DNA nanostructures, non-coding RNAs (SO. HOT.), drug design and delivery |
| - | Cell: 781 330 1969
| + | |
| | [mailto:chan.tiffany@gmail.com chan.tiffany at gmail.com] | | [mailto:chan.tiffany@gmail.com chan.tiffany at gmail.com] |
| | + | Currently taking [http://openwetware.org/wiki/Harvard:Biophysics_101/2007 Biophysics 101] |
