Sensor: Week 9 July 12-15

From 2011.igem.org

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==Tuesday, July 16==
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==Tuesday, July 12==
===Clone Cro and O<sub>R</sub>, Take 2 Results===
===Clone Cro and O<sub>R</sub>, Take 2 Results===
According to the sequencing results, the Cro and O<sub>R</sub> genes were successfully cloned into the K3 vector.
According to the sequencing results, the Cro and O<sub>R</sub> genes were successfully cloned into the K3 vector.
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The following constructs were grown up in culture in order to send to sequencing for verification:
The following constructs were grown up in culture in order to send to sequencing for verification:
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Or + K648000,
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Or + K648000
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Or + K648000 + J23113 + B0030 + C0051,
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Or + K648000 + J23113 + B0034 + C0051,
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Or + K648000 + J23113 + B0030 + C0051
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Or + K648000 + J23113 + B0034 + C0051
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Or + K648000 + J23113 + B0031 + C0051
Or + K648000 + J23113 + B0031 + C0051
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==Wednesday, July 13==
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The sensor group ordered the following oligos:
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B0030 + C0051
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B0031 + C0051
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B0034 + C0051
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The sensor group also attempted assembling two parts, one that contained both O<sub>R</sub> and the 34mCherry reporter, and one that contained Cro and the RBS B0034.
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{|border="1"
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!Protocol
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|colspan="2" align="center" | '''Part Involved with Protocol'''
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|-
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|Insert PCR
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|colspan="2"|B0034
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|-
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|rowspan="4"|Restriction Digest
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|Insert using X and P:
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|34mCherry
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|-
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|Vector using S and P:
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|O<sub>R</sub>
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|-
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|Insert using E and S:
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|B0034
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|-
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|Vector using E and X:
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|Cro
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|-
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|Ligation
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|colspan="2"|Or + 34Mcherry<br />Cro + B0034
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|-
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|Transformation/Plating
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|colspan="2"| The above ligations were transformed into<br />Escherichia coli cells and plated onto kanamycin<br />resistant plates.
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|}
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==Thursday, July 14==
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The O<sub>R</sub>+34mCherry and Cro+B0034 constructs were grown up in culture in order to extract plasmids for sequencing tomorrow.
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==Friday, July 15==
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The O<sub>R</sub>+34mCherry and Cro+B0034 plasmids were sent to sequencing, and the results were positive. Thus, freezer stocks from the cells containing these plasmids were made.
[[Team:Penn_State/Notebook| Back to Notebook]]
[[Team:Penn_State/Notebook| Back to Notebook]]

Latest revision as of 20:01, 30 July 2011

Contents

Tuesday, July 12

Clone Cro and OR, Take 2 Results

According to the sequencing results, the Cro and OR genes were successfully cloned into the K3 vector.

cI Repressor Test Assembly, Day 31

The following constructs were grown up in culture in order to send to sequencing for verification:

Or + K648000

Or + K648000 + J23113 + B0030 + C0051

Or + K648000 + J23113 + B0034 + C0051

Or + K648000 + J23113 + B0031 + C0051

Wednesday, July 13

The sensor group ordered the following oligos:

B0030 + C0051

B0031 + C0051

B0034 + C0051

The sensor group also attempted assembling two parts, one that contained both OR and the 34mCherry reporter, and one that contained Cro and the RBS B0034.

Protocol Part Involved with Protocol
Insert PCR B0034
Restriction Digest Insert using X and P: 34mCherry
Vector using S and P: OR
Insert using E and S: B0034
Vector using E and X: Cro
Ligation Or + 34Mcherry
Cro + B0034
Transformation/Plating The above ligations were transformed into
Escherichia coli cells and plated onto kanamycin
resistant plates.

Thursday, July 14

The OR+34mCherry and Cro+B0034 constructs were grown up in culture in order to extract plasmids for sequencing tomorrow.

Friday, July 15

The OR+34mCherry and Cro+B0034 plasmids were sent to sequencing, and the results were positive. Thus, freezer stocks from the cells containing these plasmids were made.

Back to Notebook

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