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From 2013.igem.org

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Revision as of 10:21, 15 August 2013

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

Ligations

We have replicates of each, so there is:

Yellow (S05058) + Terminator #1

Yellow (s05058) + Terminator #2

Magenta + Terminator #1

Magenta + Terminator #2

Yellow (K592010) + Terminator #1

Yellow (K592010) + Terminator #2

RFP + Terminator #1

RFP + Terminator #2