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