"
Team:HokkaidoU Japan/Notebook/plastic Week 8
From 2012.igem.org
(→Colony PCR) |
(→Colony PCR) |
||
| Line 171: | Line 171: | ||
==Colony PCR== | ==Colony PCR== | ||
<p> | <p> | ||
| + | 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. | ||
| + | </p> | ||
| + | ==Liquid culture== | ||
| + | <p> | ||
| + | Cultivation of bacteria holds RBS (B0034) was started. | ||
</p> | </p> | ||
| + | |||
</div></div> | </div></div> | ||
Revision as of 13:27, 31 August 2012
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
We ligated phaA and pSB1C3, phaB and pSB1C3.
Transformation
August 23th
Colony PCR
We did colony PCR of phaA and phaB. From the result, we can say that...
August 24th
Colony PCR
August 25th
Colony PCR
The length of PhaA on pSB1C3 and PhaB on pSB1C3 were confirmed by colony PCR.
The result showed PhaA and pSB1C3 were ligated correctly.
Liquid culture
Cultivation of bacteria holds RBS (B0034) was started.
August 26th
Mini prep of DNA
