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Team:HokkaidoU Japan/Notebook/plastic Week 9
From 2012.igem.org
Contents |
August 27th
Digestion
Digestion to divide BBa_K342001(PhaC) with XbaI and PstI.
And BBa_B0034(RBS) with SpeI and PstI (with three samples).
PhaC (BBa_K342001)
| DNA solution (100 ng/ul) | 12.5 ul |
| XbaI | 1 ul |
| PstI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 3.5 ul |
| Total | 20 ul |
RBS (BBa_B0034)
N0.1
| DNA solution (20.3 ng/ul) | 14.3 ul |
| SpeI | 1 ul |
| PstI | 1 ul |
| 10xH buffer | 2.5 ul |
| DW | 0.2 ul |
| Total | 25 ul |
N0.2
| DNA solution (15.6 ng/ul) | 18.6 ul |
| SpeI | 1 ul |
| PstI | 1 ul |
| 10xH buffer | 2.5 ul |
| DW | 1.9 ul |
| Total | 25 ul |
N0.3
| DNA solution (16.9 ng/ul) | 17.2 ul |
| SpeI | 1 ul |
| PstI | 1 ul |
| 10xH buffer | 2.5 ul |
| DW | 3.3 ul |
| Total | 25 ul |
Gel extraction
Gel extraction for digestion product. Used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.
August 28th
Digestion
Digestion to divide PhaA, PhaB and PhaC(BBa_K342001) with XbaI and PstI.
And PhaC(BBa_K342001) and pSB1C3 with XbaI and SpeI.
PhaA
| DNA solution (125 ng/ul) | 6.6 ul |
| XbaI | 1 ul |
| PstI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 9.4 ul |
| Total | 20 ul |
PhaB
| DNA solution (125 ng/ul) | 4 ul |
| XbaI | 1 ul |
| PstI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 12 ul |
| Total | 20 ul |
PhaC(BBa_K342001)
| DNA solution (125 ng/ul) | 10 ul |
| XbaI | 1 ul |
| PstI | 1 ul |
| 10xM buffer | 2 ul |
| DW | 0.9 ul |
| Total | 20 ul |
Gel extraction
Gel extraction for digestion product. Used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50 ul of DNA solution.
August 29th
PHB polymer ethanolysis & preparation for GC/MS
We did ethanolysis of PHB polymer for 4hrs, and did the preparation for GC/MS.
