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From 2012.igem.org

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Revision as of 15:25, 29 June 2012

Bacterial Transformation

Bacteria: T10 DNA: pCS2 (+) at 640 ng.uL-1

1) Keep constantly the cells on ice 2) Add 1 uL of pCS2 (+) in 100 uL of T10 3) Incubate 30 min on ice 4) Heat shock the cells during 30 sec at 42 degree Celsius in a water-bath without shaking 5) Put 2 min on ice 6) Add 500 uL of pre warmed SOC-medium 7) Incubate 1h at 37 degree Celsius at 225 rpm 8) Spin at 5000 rpm during 30 sec 9) Remove 150 uL - 400 uL of supernatant 10) Resuspend the pellet in the 150 uL left 11) Spread on appropriate plates 12) Incubate overnight at 37 degree Celsius

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List of promoters received:

p2 Flk-1 p2 Xlurp p2 foxi short p2 foxi long p2 pax3 p2 Ef1a p2 Rosa26 p2 LMO2 short p2 LMO2 long p2 vast p2 HB9 p2 NBT p2 HSP70 p2 pax6 p2 CMV p2 brachyury p2 vimentin p2 14 xUAS E1b-ATG