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From 2010.igem.org

Parts Assembly

Miniprep of the overnight cultures of the following parts was performed:

52b,52c 53a,53c 54a,54b,54c, 55b,55c

The subparts 6e+14f, 6e+21f were restriction digested and ligated into chloromphenicol backbone and plated for overnight growth of colonies.

The ordered construct was chemically transformed and plated. Also, it was restriction digested followed by gel purification to be assembled with part 50. The ligation was done at room temperature for 1hr 15 min followed by heat inactivation for 15 min. The ligation mix was chemically transformed, plated and kept in 37 degree room for growth of colonies.

Fusion Protein

Miniprep of the overnight cultures of MBP in backbone.

Colony PCR for the MBP took place and 3 samples was sent to sequencing.

Our ordered gene sequence arrived, so restriction digest for the required zz-luxI was done followed by gel purification. Ligation of luxI-zz into the vector and transformation chemically took place followed by plating them overnight.

More colonies were picked from the transformed ordered gene sequence and mini prep was done to obtain more of it.

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