Team:Warsaw/Calendar-Main/11 July 2008
From 2008.igem.org
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1. PCR A in 50 µl<br> | 1. PCR A in 50 µl<br> | ||
template DNA - pKS-A4 1 µl<br> | template DNA - pKS-A4 1 µl<br> | ||
- | primer | + | primer <html> |
- | primer | + | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AP+NotI_N">AP+NotI_N</a> - 2 µl<br> |
+ | primer<html> | ||
+ | <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AL+link10+homo2_N">AL+link10+homo2_N</a> - 2 µl<br> | ||
Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 µl<br> | Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 µl<br> | ||
dNTPs - 1 µl <br> | dNTPs - 1 µl <br> |
Revision as of 17:08, 26 September 2008
Preparation of constructs with OmpA protein fusions Preparation of construct omega-A template DNA - pUC19 1 µl primer OmegaLS - 2 µl primer AOmegaPli - 2 µl Pfu polymerase buffer + Mg2+ (from Fermentas) - 5 µl dNTPs - 1 µl Pfu turbo - 0.5 µl H2o - 38.5 µl program: 1. 95°C 3' 2. 95°C 30" 3. 62°C 45" 4. 72°C 45" 5. 72°C 10' 6. keeping in 4°C 25 cycles 3. gel electrophoresis 4.reisolation from agarose gel {{WarNotebookEnd}} |