Template:Team:KULeuven/26 August 2009/VanillinProduction

From 2009.igem.org

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!| Part || concentration (ng/μl) || 260/280 λ
!| Part || concentration (ng/μl) || 260/280 λ
|- align="center"
|- align="center"
-
| SamS I ||53,9 || 1,87 ||
+
| SamS I ||53,9 || 1,87  
|- align="center"
|- align="center"
-
| SamS II ||39,4 || 2,16 ||
+
| SamS II ||39,4 || 2,16
|- align="center"
|- align="center"
-
| SamS III ||57,1 || 2,07 ||
+
| SamS III ||57,1 || 2,07  
|- align="center"
|- align="center"
-
| EF I ||76,8 || 1,94 ||
+
| EF I ||76,8 || 1,94
|- align="center"
|- align="center"
-
| EF II ||83,4 || 2,04 ||
+
| EF II ||83,4 || 2,04
|- align="center"
|- align="center"
-
| EF III ||61,5 || 2,01 ||
+
| EF III ||61,5 || 2,01
|-
|-
|}
|}
* Restriction:  
* Restriction:  
-
-We performed 2 simultaneous digests: one to verify the length of the product (Restriction2Test) using SAMS I and EF III and another to prepare for ligation (Restriction4Real) with promotor and terminator using SAMS III and EF II
+
-We performed 2 simultaneous digests: one to verify the length of the product (Restriction2Test) using SAMS II and EF III and another to prepare for ligation (Restriction4Real) with promotor and terminator using SAMS III and EF II
 +
 
 +
-We cut with the 2 enzymes separately, allowing a one hour incubation in between.
 +
EF: E/S
 +
SAMS: E/S
 +
 
 +
-Restriction volumes:
 +
 
 +
[[Restriction2Test]]
 +
 
 +
{| border ="1" align="center"
 +
!| Part || DNA µl || MQ µl
 +
|- align="center"
 +
| SamS II ||12,7 || 13,3
 +
|- align="center"
 +
| EF III ||8,1 || 17,9
 +
|-
 +
|}
 +
 
 +
[[Restriction4Real]]
 +
 
 +
{| border ="1" align="center"
 +
!| Part || DNA µl || MQ µl
 +
|- align="center"
 +
| SamS III ||8,7 || 17,3
 +
|- align="center"
 +
| EF II ||6,0 || 21,0
 +
|-
 +
|}
 +
 
-
-We cut with the 2 enzymes separately, allowing a one hour incubation between
 
* Terminator was also digested
* Terminator was also digested
 +
Terminator: E/X
 +
 +
{| border ="1" align="center"
 +
!| Part || DNA µl || MQ µl
 +
|- align="center"
 +
| Terminator ||2,7 || 23,3
 +
|-
 +
|}
 +
 +
 +
* Restriction2Test: Agarose gel electrophoresis to validate if the test samples have the correct length. Signals appeared that could not be explained

Latest revision as of 08:40, 28 August 2009

  • Miniprep of fluid cultures

Nanodrop results:

Part concentration (ng/μl) 260/280 λ
SamS I 53,9 1,87
SamS II 39,4 2,16
SamS III 57,1 2,07
EF I 76,8 1,94
EF II 83,4 2,04
EF III 61,5 2,01
  • Restriction:

-We performed 2 simultaneous digests: one to verify the length of the product (Restriction2Test) using SAMS II and EF III and another to prepare for ligation (Restriction4Real) with promotor and terminator using SAMS III and EF II

-We cut with the 2 enzymes separately, allowing a one hour incubation in between. EF: E/S SAMS: E/S

-Restriction volumes:

Restriction2Test

Part DNA µl MQ µl
SamS II 12,7 13,3
EF III 8,1 17,9

Restriction4Real

Part DNA µl MQ µl
SamS III 8,7 17,3
EF II 6,0 21,0


  • Terminator was also digested

Terminator: E/X

Part DNA µl MQ µl
Terminator 2,7 23,3


  • Restriction2Test: Agarose gel electrophoresis to validate if the test samples have the correct length. Signals appeared that could not be explained
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