Team:Penn/Notebook/Biofilms

From 2012.igem.org

(Difference between revisions)
(6/21/12)
Line 32: Line 32:
==6/21/12==
==6/21/12==
*Ran gel of yesterday's PCR purification
*Ran gel of yesterday's PCR purification
-
[[File:PCR Purified eGFP & plsr.JPG]]
+
[[File:PCR Purified eGFP & plsr.JPG|thumb|120px|alt=SCIENCE.|Image of 2% Agarose Gel of post-purification PCR products of eGFP (.85kb) & plsr (.1kb)]]
*Obtained & transformed pET-26b
*Obtained & transformed pET-26b

Revision as of 18:25, 21 June 2012

Contents

6/6/12

  • Set up equipment
  • Autoclaved 2x 1L Sterile ddH2O
  • Autoclaved 500 mL LB Broth for scale up of luxS (Plate 3 Well 2H, pSB1A2), plsr (Plate 2 Well 14H in pSB1A2 resistance to Amp)
  • Added iGEM logo to wiki

6/7/12

  • Resuspended available biobricks luxS & plsr (resuspended in 10uL ddH2O)
  • Resuspended DNA transformed into DH5α (40uL DH5α+2uL DNA) & plated on LB plates w/ 15 uL of 100 mg/mL Amp spread on surface
  • Requested lsrR & lsrK from iGEM HQ

6/8/12

  • iGEM Wiki now split into two separate Notebooks
  • Plenty of colonies, too many to count(TMTC) grew from plsr & luxS transformation from 6/7/12
    • Selected one colony each
    • Grew in 10 mL LB+Amp (100 ug/mL)
  • Contacted researchers for V. harveyi bioassay & lysostaphin, V. harveyi must be purchased through ATCC ($300), lysostaphin obtained thru MTA.

6/9/12

  • ~2mL of LB evaporated during incubation
  • Cells spun down @ 5000 rpm for 10 min
  • Miniprepped 4x Lysis into 1 column
  • [plsr]=112 ng/uL
  • [luxS]=114 ng/uL

6/11/12-6/16/12

  • Dry lab work

6/18/12

  • Obtained & resuspended primers for eGFP & plsr

6/19/12

  • performed PCR w/ NEB standard taq kit w/ primers for eGFP & plsr
  • Tested 2 annealing temperatures, 55C and 50C
  • Ran gel w/ PCR product (2% agarose), saw clear eGFP bands, but possible .1kb obscured by loading dye.

6/20/12

  • Performed qiagen PCR purification & nanodropped (it like it's hot)

6/21/12

  • Ran gel of yesterday's PCR purification
SCIENCE.
Image of 2% Agarose Gel of post-purification PCR products of eGFP (.85kb) & plsr (.1kb)
  • Obtained & transformed pET-26b