Team:DTU-Denmark/Notebook/9 July 2013

From 2013.igem.org

(Difference between revisions)
(→PCR)
(→Extraction PCR)
Line 22: Line 22:
* Phusion polymerase: 0.5 ul (3.5 ul)
* Phusion polymerase: 0.5 ul (3.5 ul)
* H<sub>2</sub>O: 31.5 ul (220.5 ul)
* H<sub>2</sub>O: 31.5 ul (220.5 ul)
 +
 +
{| class="wikitable sortable" style="text-align: right"
 +
! Temperature (<sup>o</sup> C!! Time
 +
|-
 +
| Sodium Nitrate || 300
 +
|-
 +
| Sodium Nitrate || 30
 +
|-
 +
| Sodium Nitrate || 3
 +
|-
 +
| Sodium Nitrate || 0.3
 +
|-
 +
| Sodium Nitrite || 300 (Master)
 +
|-
 +
| Sodium Nitrite || 30
 +
|-
 +
| Sodium Nitrite || 3
 +
|-
 +
| Sodium Nitrite || 0.3
 +
|-
 +
| Sodium Nitrite || 0.03
 +
|-
 +
| Ammonium Acetate || 3000
 +
|-
 +
| Ammonium Acetate || 300
 +
|-
 +
| Ammonium Acetate || 30
 +
|-
 +
| Ammonium Acetate || 3
 +
|}
==Results==
==Results==

Revision as of 08:29, 10 July 2013

Contents

208

Main purpose

Run gel with 7 PCR samples of Nir operon from Pseudomonas aeruginosa and purified AMO, HAO and CYC from 08.07.2013.

Purification of Nir operon and Nanodrop measurement.

Set up new PCR reaction to extract Nir from Pseudomonas aeruginosa.

Who was in the lab

Ariadni,Henrike,Julia,Gosia

Procedure

Run gel

Purification of Nir DNA according to the protocol QIA purification protocol. (given in the Kit)

Extraction PCR

7 reactions

  • dNTP: 1 ul (7 ul)
  • HF buffer: 10 ul (70 ul)
  • Phusion polymerase: 0.5 ul (3.5 ul)
  • H2O: 31.5 ul (220.5 ul)
Temperature (o C Time
Sodium Nitrate 300
Sodium Nitrate 30
Sodium Nitrate 3
Sodium Nitrate 0.3
Sodium Nitrite 300 (Master)
Sodium Nitrite 30
Sodium Nitrite 3
Sodium Nitrite 0.3
Sodium Nitrite 0.03
Ammonium Acetate 3000
Ammonium Acetate 300
Ammonium Acetate 30
Ammonium Acetate 3

Results

Nanodrop measurement of Nir operon:

  • 17.98 ng/ul
  • 16.82 ng/ul
  • 8.51 ng/ul
Retrieved from "http://2013.igem.org/Team:DTU-Denmark/Notebook/9_July_2013"