Sensor: Week 2 May 23-27
From 2011.igem.org
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==Tuesday== | ==Tuesday== | ||
- | === | + | ===Testing cI Repressor, Day 2=== |
+ | The sensor group continued to build their test construct by inserting the constitutive promoters into the vector containing the cI repressor and ribosome binding site (B0034+C0051). The sensor group also inserted a ribosome binding site and color reporter (B0034+mCherry) in front of a terminator (B0015). | ||
[[File:Jamie_research.jpg|thumb|right|400px|Jamie says: "No more research. To the lab!"]] | [[File:Jamie_research.jpg|thumb|right|400px|Jamie says: "No more research. To the lab!"]] | ||
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Revision as of 20:26, 8 June 2011
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
Testing cI Repressor, Day 2
The sensor group continued to build their test construct by inserting the constitutive promoters into the vector containing the cI repressor and ribosome binding site (B0034+C0051). The sensor group also inserted a ribosome binding site and color reporter (B0034+mCherry) in front of a terminator (B0015).
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 |