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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
Elecrophoresis
Gel extraction
Gel ectraction of PhaA, PhaB, pSB1C3 digestion result.</ br> We used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.</ br>
August 22th
Ethanol precipitation
Ligation
Transformation
August 23th
Colony PCR
August 24th
Colony PCR
August 25th
Colony PCR
August 26th
Mini prep of DNA
