IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/22

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Team Allergy

Today, we are expecting our sequencing results from Genewiz. Once those arrive, we will check the sequences for both the primers and the correct allergen sequence (around 300bps). If the sequences are correct, we will go back to our plasmids and cut the intron out of the plasmid and digest the plasmids containing the sense strands with S, P.

Procedures

  1. Check Sequencing Results
  2. Digest Intron with X, P
  3. Digest Plasmids with Sense with S,P
  4. Gel Electrophoresis of Digested Plasmids (both Intron and Sense)
  5. Gel Extraction and Purification of Intron and Sense

Results

  • Sequencing Results
    • Ger S/A (all samples worked)
    • Bet 2S (#3 did not work) Bet 2A (all worked)
    • LTP S/A (worked)
    • Pal (#3 did not work)
    • PME

Team Fence

Sequencing Results

Barstar samples 1, 2, 5, and 6 were confirmed to be correct.

The LacI sequence was confirmed, however we were unable to confirm the presence of the NLS because of its position at the end of the gene where sequencing results are generally unreliable. (NNNNN) Lac1R and Lac3R were poor matches because of what GENEWIZ calls "Early Termination".

Barnase had some serious issues. We are re-growing colonies following ligation and intend to submit some new PCR product as well as the original ADDGENE plasmid for sequencing in the near future.

The Gal4 DNA binding domain samples 1 and 2 were perfect except for a SNP around 320 (in the case of GAL DBD 1) for our sequence and 223 of the biobricks sequence.


Gal4 DBD sample 3 was unsuccessful.

Arabidopsis PCR


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