"

From 2010.igem.org

Revision as of 15:51, 14 October 2010 (view source) AndreasConstantinou (Talk | contribs) (→Andreas) ← Older edit		Revision as of 13:19, 25 October 2010 (view source) NinaSchiller (Talk | contribs) (→Sequencing) Newer edit →
Line 62:		Line 62:
	|align="center"|ASB0045 780		|align="center"|ASB0045 780
	|}		|}
		+
		+	----
		+	==Nina==
		+
		+	===concentration measurement===
		+
		+	I measured the concentration of the gel cleaned up samples from yesterday by spectrophotometry.
		+
		+	[[Image:Aq16.jpg]]
		+
		+	===Ligation===
		+
		+	I ligated the gel cleaned samples.
		+
		+	[[Image:Aq17.jpg]]
		+
		+	===Transformation===
		+
		+	I transformed the ligation samples into nine BL21 tubes each with 100 ul cells. I added 6 ul of the ligations to the cells and I thawed the tubes in the first step in 15 min.
		+
		+	I hope this will save some time since I am skipping the transformation into Top 10 cells. We don't have much time left of the competition therefore I hope this will work.

---

Revision as of 13:19, 25 October 2010

Contents 1 Andreas 1.1 Plasmid prep 1.2 Sequencing 2 Nina 2.1 concentration measurement 2.2 Ligation 2.3 Transformation

Andreas

Plasmid prep

From 12/10 ON cultures

DNA concentration
Sample	Conc [ng/μl]	A260/A280
pSB1C3.IgGp	237.9	1.89
pSB1C3.bFGF	210.3	1.91
pSB1C3.ProtA	173.6	1.89
pSB1C3.yCCS	229.2	1.91
pSB1C3.SOD	193.0	1.87

Sequencing

• 15 μl plasmid DNA; 1.5 μl primer (VR)

DNA concentration
Sample	Label	Sequence code
pSB1C3.IgGp	pSB1C3.IgGp_VR	ASB0045 776
pSB1C3.bFGF	pSB1C3.bFGF_VR	ASB0045 777
pSB1C3.ProtA	pSB1C3.ProtA_VR	ASB0045 778
pSB1C3.yCCS	pSB1C3.yCCS_VR	ASB0045 779
pSB1C3.SOD	pSB1C3.SOD_VR	ASB0045 780

---

Nina

concentration measurement

I measured the concentration of the gel cleaned up samples from yesterday by spectrophotometry.

Ligation

I ligated the gel cleaned samples.

Transformation

I transformed the ligation samples into nine BL21 tubes each with 100 ul cells. I added 6 ul of the ligations to the cells and I thawed the tubes in the first step in 15 min.

I hope this will save some time since I am skipping the transformation into Top 10 cells. We don't have much time left of the competition therefore I hope this will work.