Template:Team:KULeuven/26 August 2009/VanillinProduction

From 2009.igem.org

(Difference between revisions)
Line 22: Line 22:
-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 I 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
+
-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 || 17,3 ||
 +
|- align="center"
 +
| EF III ||8,1 || 21,9 ||
 +
|-
 +
|}
 +
 
 +
[[Restriction4Real]]
 +
 
 +
{| border ="1" align="center"
 +
!| Part || DNA µl || MQ µl
 +
|- align="center"
 +
| SamS III ||8,7 || 21,3 ||
 +
|- align="center"
 +
| EF II ||6,0 || 24,0 ||
 +
|-
 +
|}
 +
 
 +
 
* 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 || 27,3 ||
 +
|-
 +
|}

Revision as of 09:36, 26 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 I 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 17,3
EF III 8,1 21,9

Restriction4Real

Part DNA µl MQ µl
SamS III 8,7 21,3
EF II 6,0 24,0


  • Terminator was also digested

Terminator: E/X

Part DNA µl MQ µl
Terminator 2,7 27,3
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