840:153g:Projects/project2/2008/09/23

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Part extraction and transformation of competent cells

We extracted 3 parts from the iGEM library, BBa_J45119, BBa_J45120 and BBa_I765001. Each part is assigned to a pair:

  • BBa_J45119: Diveena and Surbhi
  • BBa_J45120: Aaron and Matt
  • BBa_I765001: Jared and Corey

Competent cells that was prepared on 9.18.08 was transformed with 2uL of biobrick part in TE. Both strains that was prepared was utilized. The cells were later plated on LB+Amp plates and incubated at 37C overnight. We also discussed the work that we need to do for Wednesday and Thursday. If the transformation is successful and colonies are obtained, overnight cultures of colonies would be done on Wednesday so that we would be able to make glycerol stocks of the cells on Thursday. We also plan to design the primer needed to amplify the wintergreen productin device without the promoter present. The task of designing the primers would be done by Jared, Aaron and Diveena. Surbhi, Matt and Corey would conduct a gel electrophoresis on the positive colonies to verify the presence of the plasmid containing our biobrick part. From analyzing the map of the vector containing the UV promoter, we decided to use M-13 primers and pcr method to amplify and obtain the UV promoter region from the plasmid. Corey is in charge of removing the plates from the incubator on Wednesday morning to avoid breakdown of Ampicillin. Diveena, Matt and Aaron would meet in Dr. Axel's lab after their biochemistry class on Wednesday evening to prepare overnight cultures

Note: The punched paper containing the original DNA is stored in labelled tubes at -20C in Axel's lab.

Updated by: Diveena