Sensor: Week 10 July 18-22

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Contents

Monday, July 18

The sensor group amplified the following constructs through the PCR protocol using the reverse primer:

B0030+C0051
B0031+C0051
B0034+C0051

Tuesday, July 19

Protocol Part Involved in Protocol
Restriction Digest Inserts using EcoRI and SpeI: B0030 + C0051 (R)
B0031 + C0051 (R)
B0034 + C0051 (R)
Or
B0034
Vectors using EcoRI and XbaI: Or34mcherry
34Cro
Tev
Ligation B0030 + Or34mcherry
B0031 + Or34mcherry
B0034 + Or34mcherry
Or + 34Cro
B0034 + Tev
Transformation/Plating The above ligations were transformed into Escherichia coli
cells and plated onto kanamycin resistant plates.

Wednesday, July 20

The sensor group amplified the colonies that started growing on Tuesday, July 19 through colony PCR. The PCR products were run on an agarose gel, which indicated that all of the assemblies worked. The colonies were grown up in cultures.

Thursday, July 21

None of the cultures from Wednesday, July 20 grew any cells. The colonies from Tuesday, July 19 were grown again in culture in order for the plasmids to be extracted and sequenced.

Protocol Part Involved with Protocol
Restriction Digest Insert using XbaI and PstI: OR+K648000
Vectors using SpeI and PstI: J23113 + B0031 + C0051
J23113 + B0034 + C0051
J23113 + B0030 + C0051
Ligation J23113 + B0030 + C0051 + Or + K648000
J23113 + B0031 + C0051 + Or + K648000
J23113 + B0034 + C0051 + Or + K648000
Transformation/Plating The above ligations were transformed into Escherichia
coli cells and plated onto kanamycin resistant plates.

Results: The above assembly attempts all yielded negative sequencing results, so the assembly process had to be repeated for the above constructs.

The following constructs were created using the ordered oligonucleotides and PCR:

B0030 + C0051
B0031 + C0051
B0034 + C0051





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