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Sensor: Week 11 July 25-29
From 2011.igem.org
Contents |
Monday, July 25
| Protocol | Part Involved with Protocol | |
|---|---|---|
| Restriction Digest | Insert using EcoRI and SpeI: | B0034 |
| Vector using EcoRI and XbaI: | tev protease | |
| Ligation | B0034+tev protease | |
| Transformation/Plating | The above ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | |
OR+K64800 and OR were both grown up in culture to be used as freezer stocks.
Tuesday, July 26
| Protocol | Part Involved with Protocol | |
|---|---|---|
| Restriction Digest | Inserts using EcoRI and SpeI: | B0031 + C0051 (R) B0030 + C0051 (R) B0034 + C0051 (R) |
| Vector using EcoRI and XbaI: | OR+K648000 | |
| Ligation | B0030 + C0051 + Or + K648000 B0031 + C0051 + Or + K648000 B0034 + C0051 + Or + K648000 | |
| Transformation/Plating | The above ligations were transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | |
The B0034+tev protease assembly from Monday, July 25 was verified on an agarose gel. The gel showed a band that corresponded to the correct size, so the colonies used for the gel were grown up in culture. The sequencing results came back positive.
Wednesday, July 27
| Protocol | Part Involved with Protocol | |
|---|---|---|
| Insert PCR | OR | |
| Restriction Digest | Insert using EcoRI and SpeI: | OR |
| Vector using EcoRI and XbaI: | 34Cro | |
| Ligation | OR+34Cro | |
| Transformation/Plating | The above ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | |
The following constructs (assembled Tuesday, July 26) were verified on an agarose gel:
| B0030 + C0051 + Or + K648000 |
| B0031 + C0051 + Or + K648000 |
| B0034 + C0051 + Or + K648000 |
Results: The gel indicated that the plasmids were the correct size. The colonies used for the gel were grown up in culture.
Thursday, July 28
The constructs assembled on Tuesday, July 26, yielded poor sequencing results. Thus, the assemblies will have to be repeated.
The OR+34Cro construct assembled Wednesday, July 27, was run on an agarose gel. The gel showed that the assembly failed and will have to be repeated.
| Protocol | Part Involved with Protocol | |
|---|---|---|
| Insert PCR | B0015 | |
| Restriction Digest | Insert using XbaI and PstI: | B0015 |
| Vector using SpeI and PstI: | B0034+tev protease | |
| Ligation | B0034+tev protease+B0015 | |
| Transformation/Plating | The above ligation was transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | |
The 34Cro part was grown up in culture to be used as freezer stock.
Friday, July 29
The following plasmids were extracted using the Omega Bio-Tek miniprep kit and sent to the sequencing center:
