Team:Stockholm/20 August 2010
From 2010.igem.org
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+ | == Mimmi == | ||
+ | |||
+ | |||
+ | === pMA === | ||
+ | ==== Primer dilution ==== | ||
+ | |||
+ | {| | ||
+ | | pMA_VF | ||
+ | | width="50" | | ||
+ | | 91µl sH<sub>2</sub>O --> 100µM | ||
+ | | width="100" | | ||
+ | | 10µl + 90µl sH<sub>2</sub>O --> 10µM | ||
+ | |- | ||
+ | | pMA_VR | ||
+ | | width="50" | | ||
+ | | 91µl sH<sub>2</sub>O --> 100µM | ||
+ | | width="100" | | ||
+ | |10µl + 90µl sH<sub>2</sub>O --> 10µM | ||
+ | |} | ||
+ | |||
+ | |||
+ | |||
+ | === pMA/pEX === | ||
+ | ==== verification PCR ==== | ||
+ | |||
+ | {| | ||
+ | | | ||
+ | | | ||
+ | ! pEX | ||
+ | ! pMA | ||
+ | | colspan="6" | | ||
+ | |- | ||
+ | ! Mix | ||
+ | | (µl) | ||
+ | | X6 | ||
+ | | X3 | ||
+ | | rowspan="9" width="150" | | ||
+ | ! primers | ||
+ | | rowspan="9" width="150" | | ||
+ | ! colspan="2" | conditions | ||
+ | | rowspan="3" | | ||
+ | |- | ||
+ | | sH<sub>2</sub>O | ||
+ | | 22.5 | ||
+ | | 135 | ||
+ | | 67.5 | ||
+ | | pEX_VF | ||
+ | ! time | ||
+ | ! °C | ||
+ | |- | ||
+ | | F primer | ||
+ | | 1 | ||
+ | | 6 | ||
+ | | 3 | ||
+ | | pEX_VR | ||
+ | | 2m | ||
+ | | 94 | ||
+ | |- | ||
+ | | R primer | ||
+ | | 1 | ||
+ | | 6 | ||
+ | | 3 | ||
+ | | pMA_VF | ||
+ | | 30s | ||
+ | | 94 | ||
+ | | ) | ||
+ | |- | ||
+ | | DNA | ||
+ | | 0.5 | ||
+ | | 5X0.5 | ||
+ | | 2X0.5 | ||
+ | | pMA_VR | ||
+ | | 30s | ||
+ | | 60 | ||
+ | | > 30 cycles | ||
+ | |- | ||
+ | | align="right" | tot | ||
+ | | 25µl | ||
+ | | 250µl | ||
+ | | 75µl | ||
+ | | rowspan="3" | | ||
+ | | 2m30s | ||
+ | | 72 | ||
+ | | ) | ||
+ | |- | ||
+ | | rowspan="2" colspan="4" | | ||
+ | | 10m | ||
+ | | 72 | ||
+ | | rowspan="2" | | ||
+ | |- | ||
+ | | oo | ||
+ | | 10 | ||
+ | |} |
Revision as of 12:08, 20 August 2010
Contents |
Hassan
midnight update, after a fast recovery from sickness!
Mimmi
pMA
Primer dilution
pMA_VF | 91µl sH2O --> 100µM | 10µl + 90µl sH2O --> 10µM | ||
pMA_VR | 91µl sH2O --> 100µM | 10µl + 90µl sH2O --> 10µM |
pMA/pEX
verification PCR
pEX | pMA | ||||||||
---|---|---|---|---|---|---|---|---|---|
Mix | (µl) | X6 | X3 | primers | conditions | ||||
sH2O | 22.5 | 135 | 67.5 | pEX_VF | time | °C | |||
F primer | 1 | 6 | 3 | pEX_VR | 2m | 94 | |||
R primer | 1 | 6 | 3 | pMA_VF | 30s | 94 | ) | ||
DNA | 0.5 | 5X0.5 | 2X0.5 | pMA_VR | 30s | 60 | > 30 cycles | ||
tot | 25µl | 250µl | 75µl | 2m30s | 72 | ) | |||
10m | 72 | ||||||||
oo | 10 |