Team:Aachen/Labbook

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Contents

In order to detect the pathogens fast, specifically and inexpensively we are building sensor cells to detect these pathogens. These sensor cells can identify pathogens in very low concentration by responsing to specific extracellular molecules either secreted by or displayed on the pathogens. These molecules trigger a fast fluorescence response by our immobilized sensor cells which will be measured by our device. Aachen Cellock standingup.png


More information will be available soon!

May

8th

  • SOE-PCR step 2

23rd

  • we transformed some BioBricks and did a colony PCR
Aachen 14-05-23 COLONY-PCR.jpg
Colony PCR picture
todo













June

25th

  • preculture for competent NEB10β cells
  • submit samples for sequencing
  • make masterplates of transformed BioBricks
  • centrifuge and freeze overnight expression culture of J23101.E0240 and K516132

26th

  • made competent NEB10β cells - several things went wrong.. we'll see (remember: pre-cool centrifuge, always check if it's indeed spinning, frequently check OD of the culture)
  • colony PCR on the transformed clones looked awful. There were too many cells in the 10 µl reaction volume. Some seals weren't fully closed. Basically only primers and smear except for the positive control which contained a plasmid template instead of cells
  • 2x 500 ml of LB and three sterile flasks have been made

July

16th

  • plasmid preps
  • transformation of different reporter strains
  • ...

sample table:

BioBrick Colonies after Transformation Colony PCR result
E0240 5 positive
E0040 6 negative


22nd

  • Philipp and Michael made 100x stocks for Hartmans media
  • Michael and Vera made about 25 HM+C-plates without Glucose
  • K731520 iLOV was plated on a HM+C plate

23rd

  • Michael made 25 new HM+C plates with 1 % agar and 4 g/L glucose
  • He also added 170 µl of the Glucose stock (500 g/L) to the Glucose-free HM+C-plates
  • 1 L of sterile HM+Glucose was prepared
  • K731520 iLOV was plated on HM+C+Glucose and HM+C+Glucose drops, 3x 5 ml HM+C+Glucose precultures were inoculated (17:00)

24th

  • K731520 iLOV did not grow, neither on the plates, nor on in the liquid media. The most probable cause is that the E. coli is missing some vitamins
  • based upon the M9 recipes by the Knight and Endy labs on OpenWetWare, we made a 20 ml supplement stock solution that can be added to 1x HM media
Component supplement for 1x HM [g/L] final 1x concentration
Casamino acids 2 2 g/L
Thiamine hydrochloride 0.3 300 mg/L
$\MgSO_4$ 0.208 2 mmol/L
$\CaCl_2$ 0.011 0.1 mmol/L

The powders were dissolved in a 10 % w/v Casamino acid stock solution and sterile-filtered (.22 µm PES). 20 ml of the supplement stock can be added to 1 L of 1x HM media. To make agar chips, we can add 1 ml to the hot agar mixture, together with the three 500 µl 100x stocks for the HM.


August

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September

...

October

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