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Sensor: July 30 - September 27
From 2011.igem.org
8/1 - Digest: o OR (E +S) o 34 Cro (E +X) - Grow up: o 34 TEV + B0015 o K3 - Insert PCR: o OR + 34Cro - PCR: o C0051 + B0030 (R) o C0051 + B0031 (R) o C0051 + B0034 (R) 0 1 4 - Primers used: o 0A o 1A o 4A 8/2 - Digest: o Or+34 Cro -> E + S o 34 EV + B0015 (vector) -> E + X o K3 (vector) -> X + P o B0030 + C0051 -> X + P o B0031 + C0051 -> X + P o B0034 + C0051 -> X + P o Or + K648000 -> E + X (vector) o B0030 + C0051 -> E + S o B0031 + C0051 -> E + S o B0034 + C0051 -> E + S - Ligated and Plated: o B0030+C0051+K3 o B0031+C0051+K3 o B0034+C0051+K3 o Or+34 Cro+34 TEV+B0015 (A) o Or+34 Cro+34 TEV+B0015 (B) o B0030+C0051+Or+K648000 o B0031+C0051+Or+K648000 o B0034+C0051+Or+K648000 8/3 - Colony PCR: o B0030+C0051+K3 o B0031+C0051+K3 o B0034+C0051+K3 o Or+34 Cro+34 TEV+B0015 (A) o Or+34 Cro+34 TEV+B0015 (B) o B0030+C0051+Or+K648000 o B0031+C0051+Or+K648000 o B0034+C0051+Or+K648000 o B0030+C0051+K3 8/4 - Minipepped an sent to Sequencing: o B0034+C0051+Or+K648000 (R, F) o B0030+C0051+K3 (B) o B0030+C0051+K3 o B0030+C0051+Or+K648000 (F, R) o B0030+C0051+Or+K648000 o B0034+C0051+K3 o Or+34 Cro+34 TEV+B0015 (B) o B0021+C0051+K3 o Or+34 Cro+34 TEV+B0015 (A) o Test A o Test B - Digest: o B0030+C0051 -> E + S o B0031+C0051 -> E + S o B0034+C0051 -> E + S o Or+34 Cro+34 TEV+B0015 -> (vector) E + X - Ligation: o B0030+C0051+Or+34 Cro+34 TEV+B0015 (A + B) o B0031+C0051+Or+24 Cro+34 TEV+B0015 (A + B) o B0034+C0051+Or+34 Cro+34 TEV+B0015 (A + B) - Tranform - Stock = (Done) o B0030+C0051+K3 o B0031+C0051+K3 o B0034+C0051+K3 8/5 - Digest: o Or+34 Cro -> E + S o 34 TEV + B0015 -> E + X (vector) o K3 -> X + P (vector) o B0030+C0051 -> X + P o B0031+C0051 -> X + P o B0034+C0051 -> X + P o Or + K648000 -> E + X o B0030+C0051 -> E + S o B0031+C0051 -> E + S o B0034+C0051 -> E + S - Ligation: o B0030+C0051+K3 o B0031+C0051+K3 o B0034+C0051+K3 o Or+34 Cro+34 TEV+B0015 o B0030+C0051+Or+K648000 o B0031+C0051+Or+K648000 o Or+34 Cro+34 TEV+B0015 o B0030+C0051+Or+K648000 o B0031+C0051+Or+K648000 o B0034+C0051+Or+K648000 - Transformed and Plated – all the above 8/7 - Grew colonies from all that was plated on 8/5 8/26 - Digest: (R=Reverse) o Or+34 Cro+34 TEV+B0015 -> E + X -> Phosphatase o B0030+C0051 -> E + S o B0031+C0051 -> E + S o B0034+C0051 -> E + S - Ligated, Transformed, and Plated: o B0030+C0051+Or+34 Cro+34 TEV+B0015 o B0031+C0051+Or+34 Cro+34 TEV+B0015 o B0034+C0051+Or+34 Cro+34 TEV+B0015 8/28 - Grew up: o B0030+C0051+Or+34 Cro+34 TEV+B0015 o B0034+C0051+Or+34 Cro+34 TEV+B0015 8/29 - Miniprep and Sequence: o B0030+C0051+Or+34 Cro+34 TEV+B0015 o B0034+C0051+Or+34 Cro+34 TEV+B0015 - 1:1 Molar Solution: o 660 per base 9/9 - Digest: o B0030+C0051 -> X + S o B0031+C0051 -> X + S o B0034+C0051 -> X + S - Ran a gel: o Ladder o B0030+C0051 o B0031+C0051 o Or+34 Co+34 TEV+B0015 o B0034+C0051 - Extracted the DNA from gel and nanodropped before ligating 9/16 - Miniprepped and PCR’d: o B0030+C0051 (Primers used -> 17679 + 17680) o B0031+C0051 (Primers used -> 17681) o B0034+C0051 (Primers used -> 17682) - Tecan testing Or o Or Mcherry 1.71 -> 6µL D 2 3 4 o Or K648000 1.44 -> 7µL E 2 3 4 o Or K648000 1.02 -> 10µL F 2 3 4 o B2 .40 -> 25µL o B4 .8 -> 12 µL o B6 .65 -> 15 µL o D2 .7 -> 14 µL o D4 .8 -> 12 µL o D6 .75 -> 13 µL o F2 .4 -> 11 µL o F4 .94 -> 11 µL o F6 .9 -> 11 µL
