IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/16

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Team Allergy

More arabadopsis DNA arrived today. We are going to obtain more hpRNA parts and

  • Did a second PCR/Purification of our allergen panel from Julia's arabidopsis genomic DNA. It worked!

GFP didn't show up (first 2 lanes). The plants for the extraction weren't the GFP variety so this is expected. Genomic dna pcr 2.jpg

  • gel purifying PCR products
    • Concentrations: LTPS: LTPA: GERS: GERA: BETS: BETA: BET2S: BET2A: PME: PAL:
  • Digested Xba1 Pst1
  • Sent BetS, PDK2, and GerS (3) for sequencing
  • Miniprep amiRNA

Nanodrop of Miniprepped amiRNA Concentrations

Concentrations in ng/μL of colonies 1~6
1 2 3 4 5 6
LTP 320.7 359.4 290.2 238.5 N/A 212.4
Bet 156.8 267.7 340.0 287.4 278.9 273.5
LTP 0.5 296.4 268.8 244.2 212.0 292.6 208.3

Diagnostic Gel: Ladder, Bet1-6, LTP1-LTP4, Ladder; Ladder, LTP6, LTP.5 (1-6), Ladder

AmiRNA d digest.jpg

Team Fence

PCR of Arabidopsis Genomic DNA, attempt 2

716pcrnewdnafail.jpeg PCR failure - no product.

PCR using the newly extracted DNA was unsuccessful.

The degradation signal from the Austrians was successfully inoculated and colonies were found on the agar plates. Individual colonies from the plates were inoculated into tubes of 5 ml LB+Amp and placed in the shaking incubator overnight. (37°C)

Making Glyerol Stocks

Combined .5mL 80% glycerol with .5mL overnight cell culture of: Barnase 3 (number changed to Barnase 1 from miniprep onward), LacIN1, and Gal2.

Team Flavor

  • Did minipreps of the Mira/Brazz + Strep constructs. All constructs had failed to ligate.


  • Ran PCR of Valencene gene from extracted genomic DNA. Followed with PCR purification
  • Re-digested STOP+V0120 BioBrick, gel purified. Gel Purification failed, no DNA was extracted.