Team:DTU-Denmark/Notebook/10 July 2013

From 2013.igem.org

(Difference between revisions)
(→Comments)
(→Transformation of Biobricks)
Line 23: Line 23:
* step 6: 90 sec (instead of 60 s)at 42 <sup>o</sup>C  
* step 6: 90 sec (instead of 60 s)at 42 <sup>o</sup>C  
* step 9: Incubation without shaking
* step 9: Incubation without shaking
 +
 +
Transformation list
 +
 +
{| class="wikitable sortable" style="text-align: right"
 +
! BioBrick number!! Backbone!! Resistance!! Plate!! Well!! Copies
 +
|-
 +
| 98 || 2:00 || 1
 +
|-
 +
| 98 || 0:20 || 35
 +
|-
 +
| 66 || 0:45 || 35
 +
|-
 +
| 72 || 2:00/3:00/4:00 || 35
 +
|-
 +
| 72 || 5:00 || 1
 +
|-
 +
| 4|| ∞ || -
 +
|}
===Extraction PCR===
===Extraction PCR===

Revision as of 16:32, 10 July 2013

Contents

208

Main purpose

Run gel with 9 PCR samples of Nir operon from Pseudomonas aeruginosa

Transformation of Biobricks

PCR reaction for Nir operon

Who was in the lab

Ariadni,Henrike,Julia,Natalia

Procedure

Run gel

  • 6 samples from PCR of 09.07.2013
  • ladder broadband
  • 3 samples from purification of 09.07.2013

Transformation of Biobricks

According to the iGEM protocol ([http://parts.igem.org/Help:Protocols/Transformation transformation protocol]) with slight changes.

  • step 1: 100 μl cells
  • step 2: 1.5 μl of the resuspended DNA
  • step 6: 90 sec (instead of 60 s)at 42 oC
  • step 9: Incubation without shaking

Transformation list

BioBrick number Backbone Resistance Plate Well Copies
98 2:00 1
98 0:20 35
66 0:45 35
72 2:00/3:00/4:00 35
72 5:00 1
4 ∞ -

Extraction PCR

9 samples from culture pAO1 10 reactions

  • dNTP: 1 ul (10 ul)
  • HF buffer: 10 ul (100 ul)
  • Phusion polymerase: 0.5 ul (5 ul)
  • H2O: 31.5 ul (315 ul)

Settings for PCR with three different elongation times

Temperature (oC) Time (min) Rounds
98 2:00 1
98 0:20 35
66 0:45 35
72 2:00/3:00/4:00 35
72 5:00 1
4 ∞ -

Results

Comments

Kanamycin plates were of bad quality

Tetracyclin plates were made by adding 22.5 uL of tetracyclin stock to LB plates

  • Stock tetracyclin: 10 mg/mL
  • Final concentration in plates: 15 ug/mL
Retrieved from "http://2013.igem.org/Team:DTU-Denmark/Notebook/10_July_2013"