Team:UCopenhagen/Notebook

N O T E B O O K

• Week 1
• Week 2
• Week 3
• Week 4
• Week 5
• Week 6
• Week 7
• Week 8
• Week 9
• Week 10
• Week 11
• Week 12
• Week 13
• Week 14
• Week 15
• Week 16
• Week 17
• Week 18
• Week 19

Week 1 (June 26 - July 2)

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  Wet Lab Overview

  Ordered primers, checked cell stocks and prepared LB plates

Week 2 (July 3 - July 9)

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• 

  Wet Lab Overview

  Making plates, initial transformations, ordered more primers.

  

  Interdependency

  Made glycerol stock of MG1655. Designed primers: yddG +/- his-tag, and pointmutations in trpE and aroG

  

  Number Control

  Text

  

  Protein Import

  Inoculation and creation of glycerol stocks and streaking on LB plates of 8 bacterial cultures: Micrococcus luteus, Staphylococcus epidermis, Bacillus Cereus, Bacillus licheniformis, Serratia marcescens, Pseudomonas putida and Rhizobium meliloti. Preparation of additional LB plates without antibiotica. Creation of competent Mach1 cells and fast transformation with pRSET A to test efficiency.

Week 3 (July 10 - July 16)

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• 

  Wet Lab Overview

  Started making plates and initial transformations

  

  Interdependency

  PCR from MG1655 stock. Amplified aroG #1 and #2 in first attempt, and trpE #2 at a higher annealing temp, then began optimizing the PCR for yddG and trpE#2 using gradient temperatures without success.

  

  Number Control

  Text

  

  Protein Import

  Fluorescent microscopy of S. marascens, E. coli and S. epidermis stained with bodipu. Removal of XbaI sites in pRSET A mod, PCR amplification and gel-extraction of the plasmid. Transformation of competent cells with pRSET amod 1, and test of XbaI site removal by digestion. Started work on pRSETjin and pRSETjin CPP vectors from the pRSET vector without XbaI site.