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Minnesota/4 July 2008
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|1. '''PCR (Polymerase Chain Reaction):''' PCR used to modify Lambda_cI by copying a section of the gene, adding RBS, and using RS (restriction site enzyme) to cut copied gene section at the appropriate restriction site. | |1. '''PCR (Polymerase Chain Reaction):''' PCR used to modify Lambda_cI by copying a section of the gene, adding RBS, and using RS (restriction site enzyme) to cut copied gene section at the appropriate restriction site. | ||
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| + | |2. '''Run PCR in Gel Electrophoresis:''' | ||
| + | |- | ||
| + | |a. Used 0.8% agarose gel, TAE buffer, and ethidium bromide (interculating agent) | ||
| + | |- | ||
| + | |b. Gel proved have correct Lambda_cI gene with 750 base pairs. Can now perform PCR purification (add buffers). | ||
| + | |- | ||
| + | |3. '''Miniprep MCherry gene''' | ||
| + | |- | ||
| + | |4. Spectrophotometry: | ||
Revision as of 15:50, 7 July 2008
| 1. PCR (Polymerase Chain Reaction): PCR used to modify Lambda_cI by copying a section of the gene, adding RBS, and using RS (restriction site enzyme) to cut copied gene section at the appropriate restriction site. |
| Components | Volume |
|---|---|
| Phusion HF Buffer | 10.0uL |
| 10 mM dNTP's | 1.0uL |
| Primer mix | 2.5uL |
| Template DNA | 1.0uL |
| Phusion DNA Polymerase | 0.5uL |
| H20 | 35.0uL |
| 2. Run PCR in Gel Electrophoresis: |
| a. Used 0.8% agarose gel, TAE buffer, and ethidium bromide (interculating agent) |
| b. Gel proved have correct Lambda_cI gene with 750 base pairs. Can now perform PCR purification (add buffers). |
| 3. Miniprep MCherry gene |
| 4. Spectrophotometry: |
