IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/07/27

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(Team Allergy)
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Revision as of 11:50, 27 July 2010

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Team Allergy

  • Diagnostic Digest (EcoR1 & Pst1)

1-6: LTP + PDK (3200 & 1200);

7-16: LTP + PAL (3200 & 540);

17,18: Bet1 + PDK (3200 & 1200);

19-22: Bet1 + PAL (3200 & 540);

23,24: Bet2+PDK (3200 & 1200);

25-28: Bet2 + PAL (3200 & 540);

29-32: Ger3 + PDK (3200 & 1200);

33-38: Ger3 + PAL (3200 & 540);

Image: Ladder (every 6)

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  • Two LTPS+Pal worked (9,11), 3 Ger3S+Pal worked (36-38), 4 Bet2S+Pal Worked (25-28)
    • Digested these parts with spe and pst1 (slow digest)

Antisense-Intron Backbone Digestion

1-39
DNA ~1.5 ug
10x BSA 3
diH2O Fill to 30
Spe 1.5
Pst1 1.5
Neb Buffer 2 (10x) 3

(Let it go for 2 hours and then phosphatized backbone (20 min)

  • Ligated with antisense parts and transformed



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