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Team:HokkaidoU Japan/Notebook/plastic Week 9
From 2012.igem.org
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==Digestion== | ==Digestion== | ||
<p> | <p> | ||
| - | Digestion to divide PhaA | + | Digestion to divide PhaA and PhaB with XbaI and PstI.<br />I digested PhaC(BBa_K342001) with these restriction sites and also XhoI to divide pSB1C3 into pieces, on which PhaC is.<br />Because the length of pSB1C3 is nearly PhaC.<br />And I digested PhaC(BBa_K342001) and pSB1C3 with XbaI and SpeI.<br /><br /> |
| - | <br /><br /> | + | |
PhaA | PhaA | ||
{|class="hokkaidou-table-digestion" | {|class="hokkaidou-table-digestion" | ||
| Line 176: | Line 175: | ||
|PstI | |PstI | ||
|1 ul | |1 ul | ||
| + | |- | ||
| + | |XhoI | ||
| + | |5.1 ul | ||
|- | |- | ||
|10xM buffer | |10xM buffer | ||
Revision as of 08:40, 29 August 2012
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 and PhaB with XbaI and PstI.
I digested PhaC(BBa_K342001) with these restriction sites and also XhoI to divide pSB1C3 into pieces, on which PhaC is.
Because the length of pSB1C3 is nearly PhaC.
And I digested 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 |
| XhoI | 5.1 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
We did ethanolysis of PHB polymer for 4hrs with sample 2~8
Preparation for GC/MS
We did the preparation for GC/MS with sample 2~8.
