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From 2010.igem.org

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

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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.