Team:Heidelberg/Templates/DelH week23

From 2013.igem.org

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== 30-09 - 06-10-13 ==
== 30-09 - 06-10-13 ==
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===Generation of DelH Plasmid pHM04 15-09===
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====Colony-PCR Conditions CP.W23.A====
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{| class="wikitable" style="float:left; margin-right:1em"
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|-
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| Template || 23x 1 picked colony, positive control
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|-
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| Expected length [bp] ||  663
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|-
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| Named || 1A - 12H
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|-
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| Primer fw 10 µM ||100x  2 µl VF2 
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|-
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| Primer rev 10 µM ||100x  2 µl DN07
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|-
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| iTaq Polymerase (2x) ||100x 10 µl
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|-
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| ddH<sub>2</sub>O ||96x 6 µl 
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|}
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{| class="wikitable" style="float:left; margin-right:1em"
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|-
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! Cycles !! Temperature DelH [°C] !! Time [s]
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|-
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| 1 || 95 || 120
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|-
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| rowspan="3" | 12 || 95 || 30
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|-
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| 68 (touchdown -0.5°C) || 30
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|-
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| 72 || 45
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|-
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| rowspan="3" | 18 ||  95 || 30
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|-
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| 65  || 30
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|-
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| 72 || 45
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|-
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| 1  || 12 || inf
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|}
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====Result====
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Expected band: 663 bp
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<br/>
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[[|200px|thumb|right|'''Fig.21.1''' Colony PCR of Gibson assembled DelH-BB without mRFP 23 colonies]]
<br/>
<br/>

Revision as of 13:27, 22 October 2013

Contents

30-09 - 06-10-13

Generation of DelH Plasmid pHM04 15-09

Colony-PCR Conditions CP.W23.A

Template 23x 1 picked colony, positive control
Expected length [bp] 663
Named 1A - 12H
Primer fw 10 µM 100x 2 µl VF2
Primer rev 10 µM 100x 2 µl DN07
iTaq Polymerase (2x) 100x 10 µl
ddH2O 96x 6 µl
Cycles Temperature DelH [°C] Time [s]
1 95 120
12 95 30
68 (touchdown -0.5°C) 30
72 45
18 95 30
65 30
72 45
1 12 inf

Result

Expected band: 663 bp
[[|200px|thumb|right|Fig.21.1 Colony PCR of Gibson assembled DelH-BB without mRFP 23 colonies]]

Retrieved from "http://2013.igem.org/Team:Heidelberg/Templates/DelH_week23"