Team:Paris/25 August 2009
From 2009.igem.org
Cha.olivier (Talk | contribs) (→Lab work) |
(→Lab work) |
||
(19 intermediate revisions not shown) | |||
Line 12: | Line 12: | ||
|} | |} | ||
- | + | {{Paris2009_Calendar_Link|24_August_2009|26_August_2009}} | |
- | < | + | <center> '''August 25th''' </center> |
- | + | ||
- | </ | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
<html> | <html> | ||
Line 32: | Line 21: | ||
<div id="paris_content"> | <div id="paris_content"> | ||
</html> | </html> | ||
- | |||
==Lab work== | ==Lab work== | ||
Line 41: | Line 29: | ||
- | Digestion of OmpA | + | Digestion of OmpA Linker by SacI and Spe I |
Digestion of Ompa Signal by the same enzymes | Digestion of Ompa Signal by the same enzymes | ||
- | + | ||
+ | <center> [[Image:KR000778.PNG]] </center> | ||
+ | </div> | ||
+ | |||
+ | |||
<div class="caroline"> | <div class="caroline"> | ||
Line 62: | Line 54: | ||
Ligation pSB1A3 | Ligation pSB1A3 | ||
+ | |||
+ | A41 OmpAsignal ligation topovector | ||
</div> | </div> | ||
- | < | + | <div class="vicard"> |
+ | PCR+ check + purification | ||
</div> | </div> | ||
- | <div | + | <div class="experience"> |
+ | |||
+ | |||
+ | PCR->A34 (NTER ClyA) | ||
+ | |||
+ | Check gel--> OK | ||
+ | |||
+ | Purification-->ok | ||
+ | <center> [[Image:Nter_clyA_25aout.JPG]] </center> | ||
+ | |||
</div> | </div> | ||
- | |||
- | |||
- | == | + | <div class="vicard"> |
+ | Miniprep+Sequencing | ||
+ | </div> | ||
+ | <div class="experience"> | ||
- | + | Miniprep of all colonie of nter RFP and Cter RFP | |
+ | |||
+ | Colonie DO: | ||
+ | |||
+ | Nter RFP: for colonie | ||
+ | |||
+ | -1:1,80µg/ml | ||
+ | |||
+ | (Miniprep: 3,33µL+ oligos VF2(1/50): 2,45µL+ H20: 9,22µL) | ||
+ | |||
+ | (Miniprep: 3,33µL+ oligos VR(1/50): 2,45µL+ H20: 9,22µL) | ||
+ | |||
+ | -2:1,90µg/ml | ||
+ | |||
+ | (Miniprep: 3,16µL+ oligos VF2(1/50): 2,45µL+ H20: 9,39µL) | ||
+ | |||
+ | (Miniprep: 3,16µL+ oligos VR(1/50): 2,45µL+ H20: 9,39µL) | ||
+ | |||
+ | -3:1,65µg/ml | ||
+ | |||
+ | (Miniprep: 3,64µL+ oligos VF2(1/50): 2,45µL+ H20: 8,81µL) | ||
+ | |||
+ | |||
+ | (Miniprep: 3,64µL+ oligos VR(1/50): 2,45µL+ H20: 8,81µL) | ||
+ | |||
+ | -4:1,66µg/ml | ||
+ | |||
+ | |||
+ | (Miniprep: 3,61µL+ oligos VF2(1/50): 2,45µL+ H20: 8,94µL) | ||
+ | |||
+ | (Miniprep: 3,61µL+ oligos VR(1/50): 2,45µL+ H20: 8,94µL) | ||
+ | |||
+ | Cter RFP: for colonie | ||
+ | |||
+ | -1:1,80µg/ml | ||
+ | |||
+ | -2:1,90µg/ml | ||
+ | |||
+ | -3:1,70µg/ml | ||
+ | |||
+ | -4:1,60µg/ml | ||
+ | |||
+ | </div> |
Latest revision as of 23:20, 16 October 2009
NoteBook
|
|
|
|
|
---|
Lab work
Digestion/Gel purification w/ Soufifi
A41 OmpAsignal
PCR OmpAsignal A41 ; 70,4° ; 9s ; matrice:pINK2
Gel purif
A41 OmpAsignal A40 TolRII
Digestion X/P
Ligation pSB1A3
A41 OmpAsignal ligation topovector
PCR+ check + purification
Miniprep+Sequencing
Miniprep of all colonie of nter RFP and Cter RFP
Colonie DO:
Nter RFP: for colonie
-1:1,80µg/ml
(Miniprep: 3,33µL+ oligos VF2(1/50): 2,45µL+ H20: 9,22µL)
(Miniprep: 3,33µL+ oligos VR(1/50): 2,45µL+ H20: 9,22µL)
-2:1,90µg/ml
(Miniprep: 3,16µL+ oligos VF2(1/50): 2,45µL+ H20: 9,39µL)
(Miniprep: 3,16µL+ oligos VR(1/50): 2,45µL+ H20: 9,39µL)
-3:1,65µg/ml
(Miniprep: 3,64µL+ oligos VF2(1/50): 2,45µL+ H20: 8,81µL)
(Miniprep: 3,64µL+ oligos VR(1/50): 2,45µL+ H20: 8,81µL)
-4:1,66µg/ml
(Miniprep: 3,61µL+ oligos VF2(1/50): 2,45µL+ H20: 8,94µL)
(Miniprep: 3,61µL+ oligos VR(1/50): 2,45µL+ H20: 8,94µL)
Cter RFP: for colonie
-1:1,80µg/ml
-2:1,90µg/ml
-3:1,70µg/ml
-4:1,60µg/ml