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Sensor: Week 9 July 12-15
From 2011.igem.org
Contents |
Tuesday, July 12
Clone Cro and OR, Take 2 Results
According to the sequencing results, the Cro and OR genes were successfully cloned into the K3 vector.
cI Repressor Test Assembly, Day 31
The following constructs were grown up in culture in order to send to sequencing for verification:
Or + K648000
Or + K648000 + J23113 + B0030 + C0051
Or + K648000 + J23113 + B0034 + C0051
Or + K648000 + J23113 + B0031 + C0051
Wednesday, July 13
The sensor group ordered the following oligos:
B0030 + C0051
B0031 + C0051
B0034 + C0051
The sensor group also attempted assembling two parts, one that contained both OR and the 34mCherry reporter, and one that contained Cro and the RBS B0034.
| Protocol | Part Involved with Protocol | |
|---|---|---|
| Insert PCR | B0034 | |
| Restriction Digest | Insert using X and P: | 34mCherry |
| Vector using S and P: | OR | |
| Insert using E and S: | B0034 | |
| Vector using E and X: | Cro | |
| Ligation | Or + 34Mcherry Cro + B0034 | |
| Transformation/Plating | The above ligations were transformed into Escherichia coli cells and plated onto kanamycin resistant plates. | |
Thursday, July 14
The OR+34mCherry and Cro+B0034 constructs were grown up in culture in order to extract plasmids for sequencing tomorrow.
Friday, July 15
The OR+34mCherry and Cro+B0034 plasmids were sent to sequencing, and the results were positive. Thus, freezer stocks from the cells containing these plasmids were made.
