Team:UCLA/Notebook/Honeybee Silk/11 May 2015

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Inoculation of 5/10 Transformation

• Transformation was good
• I picked three colonies from the 1:1 kanamycin plate and suspended each in 100uL ddH2O (labeled pET 1, 2, and 3 respectively)
• Each sample was inoculated in 3 different culture tubes with 5mL LB broth and 5uL 1000X kanamycin and was incubated at 37C.

Sequencing of Silk Biobricks

We sent in both of our constructs to Genewiz for sequencing.

• For sequencing primers, we used the iGEM VF2 and VR primers that bind to the psb1c3 backbone, as well as the sequencing primer that we designed to bind to the silk sequence (p13) see https://benchling.com/s/p7bClpzQ/edit
• The sequencing results can back and checked out for sample 1 of construct 1 (silk coding region) and samples 1 and 2 for construct 2 (regulartory elements+silk)
  • We have our first honeybee biobricks!
• For sequencing results of construct 1 see https://benchling.com/s/4XT6VRRU/edit (click on alignments on the right side of screen)
• For sequencing results of construct 2 see https://benchling.com/s/TxD8z7Kq/edit (click on alignments on the right side of screen)