Team:KULeuven/Notebook/Key/Lock/AntiKey

From 2009.igem.org

Miss Blue key lock.png



Contents

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 BBa_J23110.

Week 3: 20 July 2009 - 26 July 2009

[edit] Wednesday

Liquid culture of BBa_J23110 has been made.

[edit] Thursday

The plasmid of BBa_J23110 has been isolated from the liquid culture. The glycerol stock strain containing BBa_K145201 has been plated.

[edit] Friday

The glycerol stock strain containing BBa_K145015 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:

[edit] Tuesday

Following plasmids were purified:

  • BBa_K145015: GFP with LVA tag
    • concentration: 94,1 ng/μl
    • 260/280: 1,94
  • BBa_K145201: 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