Team:Arizona State/Notebook

From 2012.igem.org

(Difference between revisions)
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* Streak plated prepared competent cells on LB no amp plate
* Streak plated prepared competent cells on LB no amp plate
** Colonies observed
** Colonies observed
 +
 +
==June 19==
 +
* Transformation (LSE)
 +
** Transformed DNA:
 +
*** T7 promoter BBa_I712074
 +
*** Constitutive promoter BBa_J23102
 +
** Cells: DH5 alpha (donated)
 +
** Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
 +
** Controls: puc19, no DNA
 +
** Plated 2 copies of each (100 ul, 250 ul) on LB amp plates.
 +
*Made 50 LB Amp plates.
 +
 +
==June 20==
 +
* Plated negative control on LB Amp plate
 +
* Liquid cultures of T7 promoter and constitutive promoter
 +
* Transformation (LSE)
 +
** Transformed DNA:
 +
*** RBS (well 1:1H BBa_B0030)
 +
*** TetR GFP (well 2:8A Part:BBa_I13522)
 +
** Cells: DH5 alpha (donated)
 +
** Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
 +
** Controls: puc19, no DNA

Revision as of 01:32, 1 October 2012


June 2012
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July 2012
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Note from Dr. Haynes: Rohit, please transfer all of the Notebook entries from the OWW Wiki
http://openwetware.org/wiki/Haynes_Lab:Notebook/ASU_iGEM
To this page. Follow the format below. I added a couple of sections to help you get started.

June 07

  • Transformation (LSE)
    • Transformed DNA:
      • lacZ (well 4:12G, I732019)
      • p + lacO (well 1:6G, R0011)
    • Cells: neb10beta (donated)
    • Protocol from: http://www.neb.com/nebecomm/products/productc3019.asp
    • Controls: puc19, no DNA (8 plates)

June 08

  • Transformation results
    • puc19: growth
    • negative control: no growth
    • lacZ, lacO: possible small colonies
  • liquid culture in amp media (100 ug / ml):
    • no growth of lacZ, lacO
    • growth of puc19

June 12

  • DH5a Competent Cell Prep
    • Streak plated cells on LB no amp plate, let grow overnight

June 13

  • Transformation (LSE)
    • Transformed DNA:
      • lacZ (well 4:12G, I732019)
      • p + lacO (well 1:6G, R0011)
    • Cells: DH5 alpha (donated)
    • Protocol from: http://openwetware.org/wiki/Haynes:Assembly101 (30 minute transformation)
    • Controls: puc19, no DNA (8 plates)
  • Transformation (istb4, Abhi)
    • Transformed DNA:
    • Cells:
    • Protocol from:
    • Controls:
  • DH5a Chemically Competent cell prep
    • Grew 2 seed colonies from streak plate in LB no amp
    • Grew controls to test for contamination
      • Both Seed colonies grew, no contamination present

June 14

  • Competent cell prep
    • Prepared CaCl2 buffer solution and CaCl2 glycerol buffer solution
    • Grew seed colony in 400mL LB no amp

June 15

  • Competent cell prep
    • Centrifuged falcon test tubes containing liquid colonies
    • Resuspended in CaCl2 buffer solution and incubated for 15 mins
    • Centrifuged and resuspended in CaCl2 glycerol buffer solution
    • Chilled overnight

June 16

  • Competent cell prep
    • Aliquotted 200uL into test tubes
    • Stored in -80C

June 17

  • Streak plated prepared competent cells on LB no amp plate
    • Colonies observed

June 19

  • Transformation (LSE)
    • Transformed DNA:
      • T7 promoter BBa_I712074
      • Constitutive promoter BBa_J23102
    • Cells: DH5 alpha (donated)
    • Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
    • Controls: puc19, no DNA
    • Plated 2 copies of each (100 ul, 250 ul) on LB amp plates.
  • Made 50 LB Amp plates.

June 20

  • Plated negative control on LB Amp plate
  • Liquid cultures of T7 promoter and constitutive promoter
  • Transformation (LSE)
    • Transformed DNA:
      • RBS (well 1:1H BBa_B0030)
      • TetR GFP (well 2:8A Part:BBa_I13522)
    • Cells: DH5 alpha (donated)
    • Protocol from: http://tools.invitrogen.com/content/sfs/manuals/subcloningefficiencydh5alpha_man.pdf (30 minute transformation)
    • Controls: puc19, no DNA
Retrieved from "http://2012.igem.org/Team:Arizona_State/Notebook"