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Sensor: Week 2 May 23-27
From 2011.igem.org
Contents |
Monday
Testing cI Repressor, Day 1
The sensor group began the assembly of a cI repressor testing construct by amplifying the below constitutive promoters via PCR.
| Assembly Step | Parts Involved in Assembly Step |
|---|---|
| PCR | J23113 J23105 J23118 J23100 |
Tuesday
| Assembly Step | Parts Involved in Assembly Step | |
| PCR Purification | J23113 J23105 J23118 J23100 | |
| Miniprep | B0051 | |
| Restriction Digest | Inserts using EcoRI and SpeI: | J23113 J23105 J23118 J23100 B0034+mCherry |
| Vectors using EcoRI and XbaI: | B0034+C0051 B0015 | |
| Ligation | J23113 and B0034+C0051 J23105 and B0034+C0051 J23118 and B0034+C0051 J23100 and B0034+C0051 B0034+mCherry and B0015 | |
| Transformation/Plating | The above ligations were transformed into Escherichia coli cells and plated onto Amp resistant plates. | |
| PCR | B0030 B0031 B0032 B0033 | |
Wednesday
| Assembly Step | Parts Involved with Assembly Step | |
|---|---|---|
| Colony PCR | B0034+mCherry+B0015 (Kan resistance) B0034+mCherry+B0015 (Amp resistance) | |
| PCR Purification | B0030 B0031 B0032 B0033 | |
| Restriction Digest | Inserts using EcoRI and SpeI: | B0030 B0031 B0032 B0033 |
| Vector using EcoRI and XbaI: | C0051 | |
| Ligation | B0030+C0051 B0031+C0051 B0032+C0051 B0033+C0051 | |
| Transformation/Plating | The above ligations were transformed into Escherichia coli cells and plated onto Amp resistant plates. | |
Thursday
| Assembly Step | Part Involved with Assembly Step | |
|---|---|---|
| Miniprep for Sequencing | B0034+mCherry+B0015 J23113+B0034+C0051 J23105+B0034+C0051 | |
| PCR | B0034+mCherry+B0015 | |
| Restriction Digest | Insert using EcoRI and SpeI: | B0034+mCherry+B0015 |
| Vectors using EcoRI and XbaI: | J23113+B0034+C0051 J23105+B0034+C0051 | |
| Ligation | B0034+mCherry+B0015 and J23113+B0034+C0051 B0034+mCherry+B0015 and J23105+B0034+C0051 | |
| Transformation/Plating | The above ligations were transformed into Escherichia coli cells and plated onto Amp resistant plates. | |
Note: The minipreps in green font indicate the sequencing proved the assembly worked.
Friday
The following assemblies have been confirmed and named for the registry.
| Assembly | Registry Name |
|---|---|
| B0034+mCherry+B0015 J23113+B0034+C0051 J23105+B0034+C0051 | K648000 K648001 K648002 |
