Team:Paris/August 25
From 2008.igem.org
(Difference between revisions)
(→Electrophoresis) |
(→Electrophoresis) |
||
Line 264: | Line 264: | ||
='''Screening of the cloning of pFlgA-YFP Tripart (LVA+/-)'''= | ='''Screening of the cloning of pFlgA-YFP Tripart (LVA+/-)'''= | ||
- | ==Electrophoresis== | + | '''Bold text'''==Electrophoresis== |
{|border="1" style="text-align: center" | {|border="1" style="text-align: center" | ||
+ | |colspan="13"|Gel n° 1 | ||
+ | |colspan="13"|Gel n° 2 | ||
+ | |- | ||
|'''well n°''' | |'''well n°''' | ||
+ | |1 | ||
+ | |2 | ||
+ | |3 | ||
+ | |4 | ||
+ | |5 | ||
+ | |6 | ||
+ | |7 | ||
+ | |8 | ||
+ | |9 | ||
+ | |10 | ||
+ | |11 | ||
+ | |12 | ||
|1 | |1 | ||
|2 | |2 | ||
Line 291: | Line 306: | ||
| | | | ||
| | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |1kb ladder | ||
+ | | | ||
+ | | | ||
+ | |coslpan="8| | ||
|- | |- | ||
|'''expected size (pb)''' | |'''expected size (pb)''' | ||
Line 299: | Line 322: | ||
|- | |- | ||
|'''measured size''' | |'''measured size''' | ||
- | |||
| | | | ||
|style="background: #cbff7B"| | |style="background: #cbff7B"| | ||
Line 308: | Line 330: | ||
|style="background: #cbff7B"| | |style="background: #cbff7B"| | ||
|style="background: #cbff7B"| | |style="background: #cbff7B"| | ||
+ | |style="background: #cbff7B"| | ||
+ | |style="background: #cbff7B"| | ||
+ | |style="background: #cbff7B"| | ||
+ | |style="background: #cbff7B"| | ||
+ | |style="background: #cbff7B"| | ||
+ | | | ||
+ | |style="background: #cbff7B"| | ||
+ | |style="background: #cbff7B"| | ||
+ | | | ||
|} | |} | ||
[[Image:KR000.jpg|thumb|Screening of L164]] | [[Image:KR000.jpg|thumb|Screening of L164]] |
Revision as of 19:27, 26 August 2008
Construction for SynchronizationTransformation of the ligations we did yesterday
Construction of pFlgA - GFP GeneratorAim : Construction of "pFlgA-RBS-GFP-dbl ter" (pFlgA-E0240)
Results of the transformation we did the day before yesterday
=> Need to screen to know which clones we can use for the of pFlgA promotor characterization. Cloning of EnvZ*The sequencing of EnvZ* previously cloned, revealed a loss of about 300 bp. EnvZ* contains indeed an EcoRI restriction site within its sequence. So we can't use this enzyme during the cloning. Digestion
Reaction mixture
Incubation at 37°C during 2H25, and then ~20 min at 65°C Electrophoresis1% agarose gel
elution in 30 µL of buffer EB
Promoter characterization plasmidsTransformation results: ligations from August 21thTop 10 cells were used
Screening of the cloning of pFlgA-YFP Tripart (LVA+/-)Bold text==Electrophoresis==
Minipreps and glycerol stock
|