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Team:SUSTC-Shenzhen/Notebook/CRISPR/Verification-for-plasmid-PX330-MCherry-GAL4-PX330-MCherry-amplified-2014-8-9-by-restriction-digestion
From 2014.igem.org
Notebook
Elements of the endeavor.
Contents |
Verification for plasmid PX330-MCherry-GAL4,PX330-MCherry (amplified on 2014.8.9) by restriction digestion
2014/8/09 Failure, as it is.Materials
- Enzyme & Buffer: EcoRI & AgeI Buffer2.1
Methods
1) Digestion system：（unit: ul）
| DNA | EcoR I | Age I | Buffer | ddH2O | |
|---|---|---|---|---|---|
| PX330-MCherry-GAL4\PX330-MCherry | 0.5 | 1 | 1 | 1 | 6.5 |
2) Loading system：（unit: ul）
| DNA | DYE | TAE | Total volume | |
|---|---|---|---|---|
| PX330-MCherry-GAL4\PX330-MCherry | 0.5 | 2 | 9.5 | 12 |
Running conditions：130V, 30min
Results
Supposed results: Two bands: 4200, 800bp Actual results:
There was no band appearing on the position that aligned to 800bp pointed by marker, and the bands of 4200bp were also very indistinct, all of which might be caused by the low concentration of our DNA sample loaded for electrophoresis.
