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Team:KULeuven/Notebook/Key/Lock/AntiKey
From 2009.igem.org
Key / Lock / AntiKey Notebook
Week 1: 6 July 2009 - 12 July 2009
Week 2: 13 July 2009 - 19 July 2009
[edit] Thursday
Plating the glycerol stock strain containing .
Week 3: 20 July 2009 - 26 July 2009
[edit] Wednesday
Liquid culture of has been made.
[edit] Thursday
The plasmid of has been isolated from the liquid culture.
The glycerol stock strain containing has been plated.
[edit] Friday
The glycerol stock strain containing has been plated.
The key+terminator sequence, the lock sequence and the antikey+terminator sequence have all been ordered at GeneArt.
Week 4: 27 July 2009 - 2 August 2009
[edit] Monday
Following parts were put in liquid medium:
- + : GFP with LVA tag and terminators
- TetR-promoter
[edit] Tuesday
Following plasmids were purified:
- GFP with LVA tag
- concentration: 94,1 ng/μl
- 260/280: 1,94
- tetR-promoter
- concentration: 98,2 ng/μl
- 260/280: 1,89
Week 5: 3 August 2009 - 9 August 2009
Week 6: 10 August 2009 - 16 August 2009
Week 7: 17 August 2009 - 23 August 2009
Week 8: 24 August 2009 - 30 August 2009
Week 9: 31 August 2009 - 6 September 2009
Week 10: 7 September 2009 - 13 September 2009
Week 11: 14 September 2009 - 20 September 2009
Week 12: 21 September 2009 - 27 September 2009
Week 13: 28 September 2009 - 4 October 2009
[edit] Tuesday
- We finally received the lock and the antisense key from geneart.
- Electroporation and plating out the two components.
[edit] Wednesday
- Liquid culture of the two components has been made
[edit] Thursday
- Miniprepped and nanodropped
| sample | ng/µl |
|---|---|
| L1 | 93,5 |
| L2 | 81,1 |
| AK1 | 75,3 |
| AK2 | 72,8 |
- Restriction digest overnight: enzymes EcoRI and PstI
[edit] Friday
- Gel electrophoresis and purification
- Ligation of lock and antisense key in pSB1A2 (over weekend)
Week 14: 5 October 2009 - 11 October 2009
[edit] Monday
- Electroporation of the ligation
- Plating out the electroporated cells
[edit] Tuesday
- No growth visible
[edit] Wednesday
- Therefore new attempt to ligate the two components in iGEM vectors
- Restriction digest: EcoRI and PstI
[edit] Thursday
- Gel electrophoresis and purification
- Ligation (overnight): both lock and antisense key are ligated in pSB1A2 and pSB3K3
[edit] Friday
- Electroporated and plated out the ligations
[edit] Sunday
- Liquid culture (the lock and antisense key in iGEM vectors)
Week 15: 12 October 2009 - 18 October 2009
[edit] Monday
Miniprep and nanodrop
- Lock (pSB1A2): ng/µl
- Antisense key (pSB3K3): ng/µl
