Team:DTU-Denmark/Notebook/24 July 2013

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===USER reaction and transformation ===
===USER reaction and transformation ===
-
We perform USER reaction in order to create construct of plasmid pZA21 and AMO and other construct - pZA21 and HAO. We add also negative control with water instead of insert.
+
We perform USER reaction, samples are as follows:
-
Reaction is performed according to our protocol with increased amount of insert.
+
 
 +
* plasmid pZA21 and AMO  
 +
* plasmid pZA21 and HAO
 +
* negative control with water instead of insert
 +
 
 +
Reaction is performed in the same way as on [[Team:DTU-Denmark/Notebook/18_July_2013 |18-07-2013]] with increased amount of insert (up to 14 uL due to low DNA concentration).
===Restriction analysis===
===Restriction analysis===
 +
 +
From last week transformants with AMO and HAO in pZA21 we performed plasmid isolation.
 +
 +
We perform restriction analysis with EcoRI. Expected fragments are as follows:
 +
*
===Primer diluting===
===Primer diluting===
===PCR to extract Nir from Pseudosomonas with new USER primers===
===PCR to extract Nir from Pseudosomonas with new USER primers===

Revision as of 13:12, 24 July 2013

Contents

208


Main purpose


Who was in the lab


Henrike, Julia, Gosia, Krystian

Procedure


USER reaction and transformation

We perform USER reaction, samples are as follows:

  • plasmid pZA21 and AMO
  • plasmid pZA21 and HAO
  • negative control with water instead of insert

Reaction is performed in the same way as on 18-07-2013 with increased amount of insert (up to 14 uL due to low DNA concentration).

Restriction analysis

From last week transformants with AMO and HAO in pZA21 we performed plasmid isolation.

We perform restriction analysis with EcoRI. Expected fragments are as follows:

Primer diluting

PCR to extract Nir from Pseudosomonas with new USER primers

Results

Conclusion

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