Team:TU Munich/Notebook/Labjournal

From 2012.igem.org

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|0.5 µl
|0.5 µl
|Pfu Turbo DNA polymerase (2.5 U / µl)
|Pfu Turbo DNA polymerase (2.5 U / µl)
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===PCR cycling parameters===
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{|cellspacing="0" border="1"
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|'''Segment'''
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|'''Cycles'''
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|'''Temperature'''
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|'''Time'''
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|-
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|1
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|1
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|95 °C
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| 30 sec
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|2
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|15
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|95°C
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| 30 sec
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|-
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|55°C
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| 1 min
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|68°C
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Revision as of 16:02, 23 June 2012

Contents

Saturday, 23th June

Quick Change mutagenis to remove NgoMIV from pYES2

Investigator: Ingmar, Volker Aim of the experiment: Generation of an RFC 25 compatible version of the pYes2 Vector.

PCR reaction batch

volume reagent
2.5 µl 10x Pfu Ultra II buffer
4 µl Plasmid P7 pYes2_RFC25 MCS template
0.5 µl 1:10 dilution of O38 (10 pmol/µL)
0.5 µl 1:10 dilution of O39 ((10 pmol/µL)
17 µl ddH2O
0.5 µl dNTP mix
0.5 µl Pfu Turbo DNA polymerase (2.5 U / µl)

PCR cycling parameters

Segment Cycles Temperature Time
1 1 95 °C 30 sec
2 15 95°C 30 sec
55°C 1 min
68°C 1 min
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