Team:HokkaidoU Japan/Notebook/plastic Week 8
From 2012.igem.org
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The length of PhaA on pSB1C3 and PhaB on pSB1C3 were confirmed by colony PCR.<br/>The result showed PhaA and pSB1C3 were ligated correctly. | The length of PhaA on pSB1C3 and PhaB on pSB1C3 were confirmed by colony PCR.<br/>The result showed PhaA and pSB1C3 were ligated correctly. | ||
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+ | [[image:HokkaidoU120825 phaAcolonyPCR.jpg|thumb|Colony PCR result]] | ||
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Revision as of 05:11, 24 September 2012
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
August 22th
August 24th
August 25th
August 26th
Plasmid extraction
The plasmid of RBS (BBa_B0034) were extracted.
And then we got 50ul DNA solution.
Digestion
RBS (BBa_B0034) was digested with SpeI and PstI restriction sites.
And also PhaC (BBa_K342001) was digested with XbaI and PstI.
liquid culture