Sensor: Week 2 May 23-27

From 2011.igem.org

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==Tuesday==
==Tuesday==
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===Testing cI Repressor, Day 2===
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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).

Jamie says: "No more research. To the lab!"
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

Jamie says: "D'oh! No one said anything about WAITING!"
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

Finally, the oligos arrived!
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





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