IGEM:University of Illinois Urbana Champaign/2009/Notebook/Bioware 2010 Arsenic Bioremediation/2010/06/29

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6/29/10

5:30PM

Digestion of backbones

Samples: pSB1A2, pSB1C3, pSB1T3, pSB2K3 (BBa_E0033), pSB1AC3, pSB1AK3, pSB1AT3

XbaI 1 ul
SpeI 1 ul
Buffer 2 5 ul
BSA .5 ul
DNA 1 ul
H2O 41.5 ul

Incubate at 37 º C, 15 min

Inactivate at 80 º C, 20 min

PCR cleanup

add 2 ul SAP buffer, 1 ul SAP

Incubate 37 º C, 10 min

Incubate 65 º C, 15 min

Gel

175 V, 1% TAE

Lane Sample Amount
1 1 Kb Ladder 2 ul
2 pSB1A2 5 ul sample + 1 ul dye
3 pSB1C3 5 ul sample + 1 ul dye
4 pSB1T3 5 ul sample + 1 ul dye
5 pSB1K3 (E0033) 5 ul sample + 1 ul dye
6 pSB1AK3 5 ul sample + 1 ul dye
7 pSB1AC3 5 ul sample + 1 ul dye
8 pSB1AT3 5 ul sample + 1 ul dye