User:TChan

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==W 6.14.06==
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<div class="tabs-blue">
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<ul>
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<li id="current">[[TChan|Project Overview]]</li>
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<li>[[TChan/Schematics|Schematics]]</li>
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<li>[[TChan/OligoList|Oligo List]]</li>
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<li>[[TChan/Scaffold|Scaffold]]</li>
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<li>[[TChan/Images|Images]]</li>
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<li>[[TChan/iGEMNotes|iGEM Notes]]</li>
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</ul>
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</div>
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<br style="clear:both">
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<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.
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*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>---
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  E0: ---|E--|<b>X</b>---gfp---|S--|<b>P</b>---
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  hypothetical ligation: ---|E--
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1. Miniprepped the 3 BioBricks Plasmids (according to [http://openwetware.org/wiki/Miniprep/Qiagen_kit standard protocol])
 
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2. Double digested the 2 R0 (lac promoter) and 2 E0 (gfp) samples
 
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==iGEM 2006 Notebook==
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<calendar>
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name=IGEM:Harvard/2006/DNA_nanostructures/Notebook
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date=2006/07/01
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view=threemonths
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format=%name/%year-%month-%day
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weekstart=7
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</calendar>
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3. Phosphatasing of 2 R0 samples to prevent self-ligation
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<calendar>
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name=IGEM:Harvard/2006/DNA_nanostructures/Notebook
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date=2006/10/01
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view=threemonths
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format=%name/%year-%month-%day
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weekstart=7
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</calendar>
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4. Ran R0 and E0 samples on gel to separate out the gfp from the E0-plasmid
 
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5. Gel extracted R0 plasmid (~2.1 kb, linearized by digestion) and E0 gfp fragment (~0.9kb)
 
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==Tu 6.13.06==
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==Notebook==
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1. Picked Colonies of Existing BioBrick Transformants
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<calendar>
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*Picked and numbered:
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name=TChan/Notebook
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  3 colonies of E7,
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date=2006/2/01
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  2 colonies of E0,
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view=threemonths
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  2 colonies of R0.
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format=%name/%year-%month-%day
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*Grew each colony in 5mL LB + 5uL of 50mg/mL Amp (1:100 dilution).
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weekstart=7
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</calendar>
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2. Ran Gel of DNA Nanostructures
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<calendar>
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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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name=TChan/Notebook
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  [[Image:Tc_mm_pt_lh_folding_0613.jpg|thumb|left|Nanostructure Gel]]
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date=2006/5/01
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view=threemonths
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  1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold
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format=%name/%year-%month-%day
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weekstart=7
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</calendar>
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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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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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===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&degC 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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1. Past iGEM Projects Presentation: UCSF
 
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2. 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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  ?
 
==Contact Info==
==Contact Info==
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   Biochemical Sciences
   Biochemical Sciences
   Kirkland '07
   Kirkland '07
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   PRISE Summer 2006
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   Academic Interests: DNA nanostructures, non-coding RNAs (SO. HOT.), drug design and delivery
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  Cell: 781 330 1969
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   [mailto:chan.tiffany@gmail.com chan.tiffany at gmail.com]
   [mailto:chan.tiffany@gmail.com chan.tiffany at gmail.com]
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  Currently taking [http://openwetware.org/wiki/Harvard:Biophysics_101/2007 Biophysics 101]

Current revision




iGEM 2006 Notebook

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Contact Info

 Tiffany Chan
 Biochemical Sciences
 Kirkland '07
 Academic Interests: DNA nanostructures, non-coding RNAs (SO. HOT.), drug design and delivery
 chan.tiffany at gmail.com
 Currently taking Biophysics 101
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