User:TChan

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(M 6.12.06)
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==
==
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==Tu 6.13.06==
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1. Picked Colonies of Existing BioBrick Transformants
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*Picked:
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  3 colonies of E7,
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  2 colonies of E0,
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  2 colonies of R0, and numbered accordingly
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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. 
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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 (NB: Only left four lanes are ours.)
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==M 6.12.06==
==M 6.12.06==
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1. Folded DNA Nanostructures
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1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA
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<b>Goal:</b> Create nanotubes using Shawn's pre-designed oligo staples, and two controls: one without the scaffold, one without the staples
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*Three reactions were mixed according to [http://openwetware.org/wiki/Folding_DNA_nanostructures  given protocol].
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*According to [http://openwetware.org/wiki/Folding_DNA_nanostructures  protocol], 3 combinations of components were mixed and thermocycled.
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oligos + scaffold
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-oligos
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-scaffold
2. Transformed Bacteria with Existing BioBrick Plasmids
2. Transformed Bacteria with Existing BioBrick Plasmids
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<b>Goal:</b> Transform existing BioBrick-containing plasmids in order to produce large enough quantities of DNA to work with later.
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*Transformed into OneShot Top10 competent cells (Invitrogen) according to the following protocol.
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*R0010 (lac operon promoter),
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  R0010 (lac operon promoter),
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*E7104 (T4 promoter + gfp),
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  E7104 (T4 promoter + gfp),  
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*and E0241 (gfp)
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  and E0241 (gfp).
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were transformed into OneShot Top10 competent cells (Invitrogen) according to
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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.

Revision as of 15:44, 14 June 2006

==

Contents

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

2. Ran Gel of DNA Nanostructures

  • Ran 2% agarose gel of the nanostructure reaction and the 2 controls for ~40 min at 130V.
Nanostructure Gel
Nanostructure Gel
 1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold (NB: Only left four lanes are ours.)
 






M 6.12.06

1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA

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).

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&degC 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) ?

Email

chan.tiffany at gmail.com

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