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From 2013.igem.org

Contents 1 208 1.1 Main purpose 1.2 Who was in the lab 1.3 Procedure 1.3.1 Run gel 1.3.2 Purification of Nir DNA according to the protocol QIA purification protocol. (given in the Kit) 1.3.3 Extraction PCR 1.4 Results

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