IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-7-12

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Contents

Experimental Results

The plates grown from yesterday show the following:

Positive Control: Many colonies

Negative Control: No colonies

Tranformed Plate: One colony

Repeat of 7/11 Topo&Transformation

Due to only one colony showing up on the transformed plate, the experiment conducted 7/11 was repeated.

  • Same DNA from 7/11
  • 6uL per transformation of DNA/topo mix
  • 2 experimental groups, 1 positive, 1 negative
  • 15uL of Top10 cells per group, except for negative - only had 5
  • After adding SOC media to all transformants, the tubes were incubated for 63 minutes.
  • The transformation mixtures were added onto plates:
  • Ampicillin plate: Positive control
  • Kanamycin plates: Experimental groups (topo) #1, #2; Negative control
    • Note: The cell spreaders for the positive control and experimental group #1 were run through the Bunsen Burner.

Purification of second gel

We purified our second gel from 7/10 using the QiaQuick gel extraction kit. The purified DNA is currently stored in a refrigerator.

Project Goal Revision

Our new flowchart
Our new flowchart

We spoke a great deal yesterday about how best to manage the time left this summer. We reviewed our project goals and made some pretty big changes, the most notable being that reinserting the KaiABC into cyanobacteria (with k.o. wild types genes) will not be pursued. We want to work specifically on attempting to create Kai oscillation in E. coli.

Incubator status

  • Removed
    • All liquid PCC 6803 cultures from the lower shelf
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