Sensor: Week 3 May 30-June 4

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Contents

Monday

cI Repressor Test Assembly, Day 6

The sensor group ran PCR, transformed, and cultured ribosome binding sites from the registry as well as the cI repressor. The sensor group also prepared for miniprep and sequencing for tomorrow.

Assembly step Parts Ivolved with Assembly Step
PCR C0051 from kit
Transform/Culture B0030
B0032
C0051 from kit
Colony Growth (for sequencing) B0031+C0051
B0033+C0051

Tuesday

cI Repressor Test Assembly, Day 7

The sensor group performed miniprep from the cultures created 5/30/2011. Two RBS and cI repressor constructs were sent to sequencing while assembly was repeated for the other two RBS and the cI repressor since this assembly from last week failed.

Assembly Step Parts Involved with Assembly Step
Miniprep For Sequencing: B0031+C0051
B0033+C0051
For Assembly: C0051
B0030
B0032
Restriction Digest Insert using XbaI and PstI: C0051
Vector using SpeI and PstI B0030
B0032
Ligation B0030+C0051
B0032+C0051
Transformation/Plating The above ligations were transformed into
Escherichia coli cells and plated onto
Amp resistant plates.

Wednesday

cI Repressor Test Assembly, Day 8

The sensor group placed constitutive promoters in front of the RBS+cI repressor construct and attatched another RBS to the cI repressor.

Assembly Step Parts Involved in Assembly Step
Restrction Digest Inserts using XbaI and PstI: J23113
J23115
C0051
Vectors using SpeI and PstI: B0033
B0031+C0051
Ligation J23113 + B0031+C0051
J23105 + B0031+C0051
B0033 + C0051
Transformation/Plating The above ligations were transformed into
Escherichia coli cells and plated onto
Amp resistant plates.

Thursday

cI Repressor Test Assembly, Day 9

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