Team:Stockholm/20 August 2010

From 2010.igem.org

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== Mimmi ==
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=== pMA ===
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==== Primer dilution ====
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{|
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| pMA_VF
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| width="50" |
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| 91µl sH<sub>2</sub>O --> 100µM
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| width="100" |
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| 10µl + 90µl sH<sub>2</sub>O --> 10µM
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|-
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| pMA_VR
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| width="50" |
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| 91µl sH<sub>2</sub>O --> 100µM
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| width="100" |
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|10µl + 90µl sH<sub>2</sub>O --> 10µM
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|}
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=== pMA/pEX ===
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==== verification PCR ====
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{|
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|
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|
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! pEX
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! pMA
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| colspan="6" |
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|-
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! Mix
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| (µl)
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| X6
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| X3
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| rowspan="9" width="150" |
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! primers
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| rowspan="9" width="150" |
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! colspan="2" | conditions
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| rowspan="3" |
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|-
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| sH<sub>2</sub>O
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| 22.5
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| 135
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| 67.5
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| pEX_VF
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! time
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! &deg;C
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|-
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| F primer
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| 1
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| 6
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| 3
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| pEX_VR
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| 2m
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| 94
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|-
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| R primer
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| 1
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| 6
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| 3
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| pMA_VF
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| 30s
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| 94
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| )
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|-
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| DNA
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| 0.5
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| 5X0.5
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| 2X0.5
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| pMA_VR
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| 30s
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| 60
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| > 30 cycles
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|-
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| align="right" | tot
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| 25µl
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| 250µl
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| 75µl
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| rowspan="3" |
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| 2m30s
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| 72
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| )
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|-
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| rowspan="2" colspan="4" |
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| 10m
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| 72
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| rowspan="2" |
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|-
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| oo
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| 10
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|}

Revision as of 12:08, 20 August 2010

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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
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