Difference between revisions of "Team:Paris Saclay/Notebook/October/2"

(Created page with "=Sunday 2<sup>nd</sup> October= ==Lab work== ===Visualization=== ====Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation==== ''By...")
 
Line 1: Line 1:
 +
{{Team:Paris_Saclay/notebook_header}}
 +
 
=Sunday 2<sup>nd</sup> October=
 
=Sunday 2<sup>nd</sup> October=
 
==Lab work==
 
==Lab work==
Line 5: Line 7:
 
====Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation====
 
====Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation====
 
''By Maxence & Victor''
 
''By Maxence & Victor''
 
  
 
PRECIPITATION ETOH
 
PRECIPITATION ETOH
 
  
 
Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:
 
Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:

Revision as of 17:12, 2 October 2016

Sunday 2nd October

Lab work

Visualization

Cloning of FRB - GFP 11 (pPS16_019) in FKBP - GFP 10 - pSB1C3 (pPS16_018) by digestion-ligation

By Maxence & Victor

PRECIPITATION ETOH

Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:

  • 1.5 µL of Buffer T4 10X
  • 1 µL of ligase T4 enzyme
  • 1 µL of ligase T4 enzyme
  • 12.5 µL of water

The mix were incubated for 30 minutes at rooming temperature.

Cloning of GFP 1.9 from pUC19 (pPS16_009) in pSB1C3 by digestion-ligation

By Maxence & Victor'

Once template and vector were cut, DNA ligase was used to join the sticky ends of the template and vector together:

  • 6 µL of template (pPS16_009 treated by PstI & XbaI)
  • 3 µL of vector (Bba treated by PstI & XbaI)
  • 1.5 µL of Buffer T4 10X
  • 1 µL of ligase T4 enzyme
  • 3.5 µL of water

The mix were incubated for 30 minutes at rooming temperature.