Haynes Lab:Notebook/Characterizing AHL quorum sensing homologs/2013/10/21

From OpenWetWare

Jump to: navigation, search
(mm/dd/yyyy)
Current revision (21:18, 21 October 2013) (view source)
(mm/dd/yyyy)
 
Line 7: Line 7:
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==mm/dd/yyyy==
==mm/dd/yyyy==
-
Retried
+
 
 +
Retry ligation from 10/19/13 '''again''' with gel extracted J06702
 +
<br>
 +
Backbone: J06702 E/X: 44 ng/ul<br>
 +
:Cut, gel purified
 +
Insert: Gblck pLac-RBS E/S: 2.6ng/ul
 +
:Cut, column purified<br>
 +
''same calculations from before, still only actually tried 3:1 and bb ctrl''<br>
 +
{| {{table}}
 +
|-
 +
| ||1:1||2:1||3:1||Bb ctrl
 +
|-
 +
|insert||0.5||1.0||1.5||0
 +
|-
 +
|vector||1.13||1.13||1.13||1.13
 +
|-
 +
|2x Roche ligation buffer||5||5||5||5
 +
|-
 +
|T4 ligase||1||1||1||1
 +
|-
 +
|H20||2.37||1.87||1.37||2.87
 +
|-
 +
|}
 +
<br>
 +
ligations ran for 3 hours at RT
 +
<br>
 +
Incubate with 50ul DH5αT for 45 min on ice
 +
<br>
 +
heat shock for 30s at 42°C
 +
<br>
 +
ice for 2min
 +
<br>
 +
spread
 +
 
 +
<!-- ##### DO NOT edit below this line unless you know what you are doing. ##### -->
 +
|}
 +
 
 +
__NOTOC__

Current revision

Project name Main project page
Previous entry      Next entry

mm/dd/yyyy

Retry ligation from 10/19/13 again with gel extracted J06702
Backbone: J06702 E/X: 44 ng/ul

Cut, gel purified

Insert: Gblck pLac-RBS E/S: 2.6ng/ul

Cut, column purified

same calculations from before, still only actually tried 3:1 and bb ctrl

1:12:13:1Bb ctrl
insert0.51.01.50
vector1.131.131.131.13
2x Roche ligation buffer5555
T4 ligase1111
H202.371.871.372.87


ligations ran for 3 hours at RT
Incubate with 50ul DH5αT for 45 min on ice
heat shock for 30s at 42°C
ice for 2min
spread



|}


Personal tools