Melbourne/Lab BL Notebook/PCR1
From 2007.igem.org
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32.5ul ddH<sub>2</sub>O | 32.5ul ddH<sub>2</sub>O | ||
| + | |||
|94°C - 5' | |94°C - 5' | ||
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4°C forever | 4°C forever | ||
| - | + | |- | |
|Reaction A => Primer BL_Con1_as | |Reaction A => Primer BL_Con1_as | ||
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| - | + | | 50ul Total | |
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Revision as of 15:12, 10 October 2007
Protocol for PCR reactions A~G
For amplifying the photoreceptor and transmembrane domains of NpSopII-NpHtrII.
| PCR mix | PCR program | PrimerII |
|---|---|---|
| 5ul 10x buffer\\
5ul Enhancer 0.6ul dNTPs (25mM stock) 2.5ul MgSO4 (Supplied in PCR kit) 1.5ul Primer BL_FP1_s (10uM stock) 1.5ul Primer II (10uM stock) 1ul Template (pJS010) 0.4ul Pfx Platinum (Invitrogen) 32.5ul ddH2O | 94°C - 5'
94°C - 30" 59°C - 30" 68°C - 35" (goto step 2 x30) 68°C - 1.5' 4°C forever | |
| Reaction A => Primer BL_Con1_as
Reaction B => Primer BL_Con2_as Reaction C => Primer BL_Con3_as Reaction D => Primer BL_Con4_as Reaction E => Primer BL_Con5_as Reaction F => Primer BL_Con6_as Reaction G => Primer BL_Con7_as | ||
| 50ul Total |
Protocol for PCR reactions 1-7
For amplification of the kinase domain of ComP
| PCR mix | PCR program |
|---|---|
| 5ul 10x buffer\\
2.5ul MgSO4 (Supplied in PCR kit) 1.5ul Primer I (10uM stock) 1.5ul Primer VR (10uM stock) 1ul Template (pJS010) 0.4ul Pfx Platinum (Invitrogen) 32.5ul ddH2O | 94°C - 5'
94°C - 30" 59°C - 30" 68°C - 35" (goto step 2 x30) 68°C - 1.5' 4°C forever |
| 50ul Total |
