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From 2011.igem.org

Contents 1 Tuesday, July 12 1.1 Clone Cro and OR, Take 2 Results 1.2 cI Repressor Test Assembly, Day 31 2 Wednesday, July 13 3 Thursday, July 14 4 Friday, July 15

Tuesday, July 12

Clone Cro and O_R, Take 2 Results

According to the sequencing results, the Cro and O_R 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 O_R 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 O_R+34mCherry and Cro+B0034 constructs were grown up in culture in order to extract plasmids for sequencing tomorrow.

Friday, July 15

The O_R+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