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iGEM 2011
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Objectives
- Gel extraccion of PCRs
- PCR for Insert. Again! Low concentration in gel extraction.
- Quiagen Gel Extraction Kit, with elution 30μL.
- Lane 1 Ladder.
- Lane 2 Insert with Prefix and suffix Standard for Gibson
- Lane 3 Backbone with Prefix and suffix complete.
I need to repeat the insert PCR and make a new Gel extraction for that biopart.
PCR for inserts
Reagents
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PCR 1 P1A7/2011
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PCR 2 P1A7/2011
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PCR 3 P1A7/2011(Control+)
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DNA
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1μL
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1μL
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1μL
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Buffer 5x PhusionHF
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10μL
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10μL
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10μL
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dNTPs(8mM)
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5μL
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5μL
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5μL
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Oligo(Extnd_Pref_Fwd)
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5μL
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5μL
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5μL‡
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Oligo(Extnd_Suff_Rev)
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5μL
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5μL
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5μL‡
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Phusion
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.5μL
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.5μL
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.5μL
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H2O
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23.5μL
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23.5μL
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23.5μL
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‡Prefix_Fwd and Suffix_Reverse Primers.
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