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Team:HokkaidoU Japan/Notebook/plastic Week 8
From 2012.igem.org
Contents |
August 20th
Digestion
A digestion to divide PhaC and pSB1C3 with XbaI and SpeI. </ br>
| DNA solution PhaC | 12 ul |
| XbaI | 1 ul |
| SpeI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 4 ul |
| Total | 20 ul |
A digestion to divide PhaA and XbaI site and SpeI site.</ br>
| DNA solution PhaA | 7 ul |
| XbaI | 1 ul |
| SpeI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 9 ul |
| Total | 20 ul |
A digestion to divide PhaB and XbaI site and SpeI site.</ br>
| DNA solution PhaB | 7 ul |
| XbaI | 1 ul |
| SpeI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 9 ul |
| Total | 20 ul |
August 21th
Electrophoresis
Electrophoresis of digested phaC and non digested phaC. To check the PCR of phaA and phaB, we also did electrophoresis of them.
Gel extraction
Gel ectraction of PhaA, PhaB, pSB1C3 digestion result. We used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.
August 22th
Ethanol precipitation
Ligation
Transformation
August 23th
Colony PCR
August 24th
Colony PCR
August 25th
Colony PCR
August 26th
Mini prep of DNA
