"

From 2013.igem.org

Revision as of 08:32, 10 July 2013 (view source) NataliaP (Talk | contribs) (→Extraction PCR) ← Older edit		Revision as of 08:35, 10 July 2013 (view source) NataliaP (Talk | contribs) (→Extraction PCR) Newer edit →
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"		{| class="wikitable sortable" style="text-align: right"
-	! Temperature (<sup>o</sup>C)!! Time	+	! Temperature (<sup>o</sup>C)!! Time (min)!! rounds
	|-		|-
-	| 98 || 300	+	| 98 || 2:00 || 1
	|-		|-
-	| 98 || 30	+	| 98 || 0:20 || 35
	|-		|-
-	| 66 || 3	+	| 66 || 0:45 || 35
	|-		|-
-	| 72 || 0.3	+	| 72 || 9:00 || 35
	|-		|-
-	| 72 || 5 min	+	| 72 || 5:00 || 1
	|-		|-
-	| 4|| ∞	+	| 4|| ∞ || -
	|}		|}

---

Revision as of 08:35, 10 July 2013

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 (oC)	Time (min)	rounds
98	2:00	1
98	0:20	35
66	0:45	35
72	9:00	35
72	5:00	1
4	∞	-

Results

Nanodrop measurement of Nir operon:

• 17.98 ng/ul
• 16.82 ng/ul
• 8.51 ng/ul