Melbourne/Lab BL Notebook/PCR1
From 2007.igem.org
(Difference between revisions)
| Line 26: | Line 26: | ||
|94°C - 5' | |94°C - 5' | ||
| - | 94°C - 30 | + | 94°C - 30" |
| - | 59°C - 30 | + | 59°C - 30" |
| - | 68°C - 35 | + | 68°C - 35" (goto step 2 x30) |
68°C - 1.5' | 68°C - 1.5' | ||
Revision as of 15:01, 10 October 2007
Protocol for PCR reactions A~G
For amplifying the photoreceptor and transmembrane domains of NpSopII-NpHtrII.
| PCR mix | PCR program |
|---|---|
| 5ul 10x buffer\\
5ul Enhancer 0.6ul dNTPs (25mM stock) 2.5ul MgSO4 (Supplied in PCR kit) 1.5ul Primer I (10uM) 1.5ul Primer II (10uM) 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 |
|-
PCR mix
|5ul 10x buffer\\
5ul Enhancer
0.6ul dNTPs (25mM stock)
2.5ul MgSO4 (Supplied in PCR kit)
1.5ul Primer I (10uM)
1.5ul Primer II (10uM)
1ul Template (pJS010)
0.4ul Pfx Platinum (Invitrogen)
32.5ul ddH2O
---
50ul Total | |-
