IGEM:Harvard/2006/Cyanobacteria/Notebook/2006-8-3

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Contents

To-do short term

  • PCR purify the second batch of digested KaiA/B.
  • Gel purify the excised BioBrick backbone (2kb fragment)
    • Ligate the above two products
  • Run the second batch of digested RFP BioBrick plasmid on a gel Failed
    • Gel purify the insert
    • (Maybe) Gel purify the backbone
      • Add phosphatase to the backbone to make it ready for ligation
  • Prepare our mysterious 400b segment for sequencing

Sequencing

  • ZS101: LC1PUR1 w/ crossF
  • ZS102: LC1PUR1 w/ crossR
  • ZS103: LC2PUR1 w/ crossF
  • ZS104: LC2PUR2 w/ crossR
  • ZS105: 400bp (stared) w/ crossF
  • ZS106: 400bp (stared) w/ crossR
  • ZS107: 400bp (circled) w/ crossF
  • ZS108: 400bp (circled) w/crossR

Expected sequences

  • 3-10: 1-210 = 210bp
  • 9-8: 189-559 = 371bp
  • 7-6: 538-2940 = 2403bp
  • 5-4: 2922-3009 = 88bp

Nanodropped amounts (ng/ul)

  • Water: 0.9. 0.7
  • KaiA: 4.1, 4.6
  • KaiB: 1 (before vortexing), 10.5 (after vortexing)
  • BB backbone high copy 1: 10.6 (only curve that looks ok)
  • BB backbone high copy 2: 9.9
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