User:TChan: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
No edit summary
Line 1: Line 1:
==
==
==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. 
  [[Image:Tc_mm_pt_lh_folding_0613.jpg|thumb|left|Nanostructure Gel]]
 
  1kb Ladder | Oligo + Scaffold | -Oligo | -Scaffold (NB: Only left four lanes are ours.)
 


==M 6.12.06==
==M 6.12.06==


1. Folded DNA Nanostructures
1. Folded DNA Nanostructures Using Shawn's Pre-designed Oligo Staples and Scaffold ssDNA
<b>Goal:</b> Create nanotubes using Shawn's pre-designed oligo staples, and two controls: one without the scaffold, one without the staples
*Three reactions were mixed according to [http://openwetware.org/wiki/Folding_DNA_nanostructures  given protocol].
*According to [http://openwetware.org/wiki/Folding_DNA_nanostructures  protocol], 3 combinations of components were mixed and thermocycled.
oligos + scaffold
-oligos
-scaffold


2. Transformed Bacteria with Existing BioBrick Plasmids
2. Transformed Bacteria with Existing BioBrick Plasmids
<b>Goal:</b> Transform existing BioBrick-containing plasmids in order to produce large enough quantities of DNA to work with later.
*Transformed into OneShot Top10 competent cells (Invitrogen) according to the following protocol.
*R0010 (lac operon promoter),
  R0010 (lac operon promoter),
*E7104 (T4 promoter + gfp),
  E7104 (T4 promoter + gfp),  
*and E0241 (gfp)
  and E0241 (gfp).
were transformed into OneShot Top10 competent cells (Invitrogen) according to
 


===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.




Revision as of 12:44, 14 June 2006

==

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
 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

Retrieved from "https://openwetware.org/mediawiki/index.php?title=User:TChan&oldid=41744"