Team:Stockholm/21 September 2010

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(New page: {{Stockholm/Top2}} == Mimmi == === CONTROL === ==== PCR ==== {| | ''pEX.SOD'' | rowspan="4" width="50" | | ''pEX.yCCS'' | rowspan="4" width="50" | | ''pEX.SOD.his'' | rowspan="4" wi...)
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Revision as of 12:55, 4 October 2010

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Contents

Mimmi

CONTROL

PCR

pEX.SOD pEX.yCCS pEX.SOD.his pEX.his.SOD
*plasmid *plasmid 1&2 *plasmid *plasmid
*Top10 2 clones *BL21 2 clones from 1&2 *BL21 2 clones *BL21 2 clones
*BL21 2 new clones *Top10 2 clones *Top10 2 clones
pSB1C3.his.SOD.cTAT
*Top10 6 clones



conditions
mix pEX x20 pSB x7 time °C
mastermix 24.5 24.5 5m 95
DNA 0.5 0.5 30s 95 )
tot 25µl 25µl 30s 55 > 30 cycles
1m20s 72 )
5m 72
oo 10


Gel

Place for picture.jpg
well sample well sample
1 ladder 12 ladder
2 pEX.SOD.his 13 pEX.SOD
3 BL21 pEX.SOD.his 1 14 Top10 pEX.SOD 1
4 BL21 pEX.SOD.his 2 15 Top10 pEX.SOD 2
5 Top10 pEX.SOD.his 1 16 pEX.yCCS 5
6 Top10 pEX.SOD.his 2 17 pEX.yCCS 8
7 pEX.his.SOD 18 BL21 pEX.yCCS 1(5) glycerol
8 BL21 pEX.his.SOD 1 19 BL21 pEX.yCCS 2(8) glycerol
9 BL21 pEX.his.SOD 2 20 BL21 pEX.yCCS 1 new
10 Top10 pEX.his.SOD 1 21 BL21 pEX.yCCS 2 new
11 Top10 pEX.his.SOD 2 22 pEX



Place for picture.jpg
well sample
1 ladder
2 pSB1C3.his.SOD.cTAT 1
3 pSB1C3.his.SOD.cTAT 2
4 pSB1C3.his.SOD.cTAT 3
5 pSB1C3.his.SOD.cTAT 4
6 pSB1C3.his.SOD.cTAT 5
7 pSB1C3.his.SOD.cTAT 6
8 pSB1C3.his.SOD





SOD / SOD.his / his.SOD / yCCS

Re-transformation

  • Quick transformation
    • 1µl DNA into 500µl BL21 cells
    • Keep on ice
    • Heat shock in 42°C for 50s
    • Cool down on ice
    • Plate on an AMP plate
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