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Team:Colombia/Notebook/Protocols
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| - | ==''' | + | =='''Protocols'''== |
| - | ==''' | + | ==='''Bacterial DNA extraction protocol'''=== |
| - | =='''Electroporation'''== | + | ==='''Miniprep'''=== |
| + | |||
| + | ==='''Electroporation'''=== | ||
Electro-competent cells of E. coli DH5α are electroporated at 1.25 mV, immediately we add SOC medium to the bacteria and we incubated on a shaker at 37°C for one hour, then, we culture cells in a solid medium containing the antibiotic of selection. | Electro-competent cells of E. coli DH5α are electroporated at 1.25 mV, immediately we add SOC medium to the bacteria and we incubated on a shaker at 37°C for one hour, then, we culture cells in a solid medium containing the antibiotic of selection. | ||
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| - | =='''PCR - Invitrogen (R) Taq polimerase'''== | + | ==='''PCR - Invitrogen (R) Taq polimerase'''=== |
Revision as of 21:52, 13 July 2012
Contents |
Protocols
Bacterial DNA extraction protocol
Miniprep
Electroporation
Electro-competent cells of E. coli DH5α are electroporated at 1.25 mV, immediately we add SOC medium to the bacteria and we incubated on a shaker at 37°C for one hour, then, we culture cells in a solid medium containing the antibiotic of selection.
PCR - Pfu DNA polimerase
One reaction
| Reactives | Amount (μL) |
|---|---|
| H2O | 16.95 |
| Buffer | 2 |
| MgSO4 | 1.6 |
| dNTP's | 0.4 |
| Primer Fw | 0.4 |
| Primer Rv | 0.4 |
| Taq | 0.06 |
| Pfu | 0.19 |
| DNA | 2 |
| Total | 10 |
