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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**Plate on an AMP plate | **Plate on an AMP plate | ||
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Latest revision as of 11:05, 26 October 2010
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
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 |
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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