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Team:SUSTC-Shenzhen/Notebook/CRISPR/Endotoxin-Free-plasmid-purification-for-pX330-mCherry-pX330-mCherry-GAL4
From 2014.igem.org
Notebook
Elements of the endeavor.
Contents |
Endotoxin Free plasmid purification for pX330+mCherry, pX330+mCherry+GAL4
2014/8/21~2014/8/22
Materials
EndoFree Plasmid Kit (TIANGEN)
Methods
1. Picked up 2 monoclones from each plate (pX330+mCherry12ml&pX330+mCherry+GAL4)
2. Incubate:
Incubate in 18ml ampicillin medium for pX330+mCherry plasmid, and 22ml for pX330+mCherry+GAL4 plasmid. Put in shaker at 37C for 14h [8.12 6:45pm~8.22 8:45 am]
3. Endotoxin Free plasmid purification for pX330+mCherry, pX330+mCherry+GAL4
Performed as the usual protocol provided by TIANGEN plasmid Kit, except that we eluted for three times at the last step with 100ul EB solution.
Results
| Concentration（ng/ul） | A260/A280 | A260/A230 | |
|---|---|---|---|
| pX330+mCherry | 453.7 | 1.90 | 2.52 |
| pX330+mCherry+GAL4 | 644 | 1.92 | 2.60 |
