IGEM:Harvard/2009/Notebook/Harvard iGEM 2010/2010/08/06

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(Team Allergy)
(Team Flavor)
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====Analysis====
====Analysis====
Both the backbone and the insert in lane 1 look like the correct length (3.2 and 1.1 kb respectively).  Lane two consists of a backbone of the correct length, but the insert appears to be ~1.5 kb, too long for the J45004 BioBrick part.  Lane 3 appears undigested; we are uncertain as to why.
Both the backbone and the insert in lane 1 look like the correct length (3.2 and 1.1 kb respectively).  Lane two consists of a backbone of the correct length, but the insert appears to be ~1.5 kb, too long for the J45004 BioBrick part.  Lane 3 appears undigested; we are uncertain as to why.
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===Transformation of Mira/Brazz StrepII constucts into E45===
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*100-200 ng of construct DNA was transformed with 15μL of E45 expression ''E. Coli''; transformation procedure was followed; transformed cells were plated on LB+AMP and left to grow O/N
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Transformations:
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Mira N1
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Mira N2
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Mira C1
 +
Mira C2
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Brazz N1
 +
Brazz N2
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Brazz C1
 +
Brazz C2

Revision as of 14:12, 6 August 2010

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Team Allergy

amiRNA

  • amiRNA-transformed turbo cells didn't grow in YEB medium. There should be more LB+Amp available by the end of the day. In the meantime, we're digesting some more backbone (8x20ul X+P+B21)
  • ligations of Bet/LTP into V0120

ihpRNA

  • no colonies on plates from transformations done yesterday after 16 hours (we let these plates grow for longer)
  • redoing ligations of ihpRNA inserts in V9/V10
    • Redigested and gel purified some backbone

[[Image: |240px|V9 and V10 backbone]]

Concentrations: V9 (5 ng/uL); V10 (6 ng/uL)

    • 12 ligations each for V9 and V10

Team Flavor

Confirmation Digest of J45004

  • J45004 in V0120 backbone; digested ~500ng DNA with Xba1/Pst1


Note: both gels images are the same, but with different contrasts/resolutions

  1. 1 kb Ladder (Invitrogen)
  2. 004-1
  3. 004-2
  4. 004-3
  5. Ladder

Backbone: 3.2 kb; Insert: 1.1 kb

Analysis

Both the backbone and the insert in lane 1 look like the correct length (3.2 and 1.1 kb respectively). Lane two consists of a backbone of the correct length, but the insert appears to be ~1.5 kb, too long for the J45004 BioBrick part. Lane 3 appears undigested; we are uncertain as to why.

Transformation of Mira/Brazz StrepII constucts into E45

  • 100-200 ng of construct DNA was transformed with 15μL of E45 expression E. Coli; transformation procedure was followed; transformed cells were plated on LB+AMP and left to grow O/N

Transformations: Mira N1 Mira N2 Mira C1 Mira C2 Brazz N1 Brazz N2 Brazz C1 Brazz C2



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