Exeter/9 August 2013

From 2013.igem.org

Exeter iGEM 2013 · Paint by Coli

Digestions of CMY

Culture NanoDrop (ng/ul) ul to get 250ng
Magenta K592012 106.9 2.34
Yellow K592010 57.1 4.38
Yellow S05058 67.1 3.73
RFP control 42.9 5.83
AMP plasmid 82.5 3.03
B0015 52.1 4.80


Xul DNA

2.5ul NEB2

0.5ul BSA (make sure diluted)

0.5ul of each required enzyme

(make up to 16ul with no-nuclease water)


Working out water needed:


Magenta = 2.34 + x = 16 x = 13.66ul

Yellow K592010 = 4.38 + x = 16 x = 11.62ul

Yellow S05058 = 3.73 + x = 16 x = 12.27ul

RFP = 5.83 + x = 16 x = 10.17ul

AMP plasmid = 3.03 + x = 16 x = 12.97ul

B0015 = 4.80 + x = 16 x = 11.20ul


Gel

Lane number Content
1 Hyperladder 1kb
2 Yellow (S05058)
3 Magenta
4 Yellow (K592010)
5 RFP
6 AMP plasmid
7 B0015
8 Hyperladder 1kb
Image: 500 pixels

Gel has had limited success. Obviously, parts like [http://parts.igem.org/Part:BBa_B0015 B0015] are too small to see on this gel, as they run much further than the ladder can indicate. Yellow and RFP are the only clear bands on the gel.

Ligations

We have replicates of each, so there is:

Yellow ([http://parts.igem.org/Part:BBa_S05058 S05058]) + Terminator #1

Yellow ([http://parts.igem.org/Part:BBa_S05058 S05058]) + Terminator #2

Magenta + Terminator #1

Magenta + Terminator #2

Yellow ([http://parts.igem.org/Part:BBa_K592010 K592010]) + Terminator #1

Yellow ([http://parts.igem.org/Part:BBa_K592010 K592010]) + Terminator #2

RFP + Terminator #1

RFP + Terminator #2

Take me back to the notebook.

Exeter iGEM 2013 · Paint by Coli