Team:Freiburg/Notebook/19 August

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This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

Contents

Meeting

attendants: Jakob, Julia, Manuel, Rüdiger, Sandra, Sophie, Theo
Time: 9:00 - 10:00

green light receptor

already done:

  • Ccas some colonies (check if positive)
  • CcaR still doesn't work

To-do:

  • cph8 still won't work, we should ask to get the original sequence

blue light receptor

already done:

  • receptor and NOT-Gate assembly

To-do:

  • add Promotor-RBS and terminator

red light receptor

already done:

  • Promotor-RBS-ho1-terminator ready
  • Promotor-RBS-pcyA-terminator ready

To-do:

  • receptor still doesn't work (not possible to amplify via a PCR)

Lysis cassette

already done:

  • quick change with the 11 bases didn't work
  • did a 3A assembly with the single parts

To-do:


Precipitator

already done:

  • finally the genes arrived
  • GST from the bioss toolkit can't be amplified from the supported vector, Rüdiger should do a positive control next time, if it still doesn't work he should get new primer pairs

To-do:

  • PBD + inducible promotor

other stuff

  • Hauke Busch recommended CellDesigner to Rüdiger to do the modelling, Rüdiger will download it and try to install it
  • modelling should be done for the precipitator, escpecially the Ni-Histidin-binding and for the plastic binding domain, maybe also the GST-Tag affinity
  • Tobi will organize moving the stuff out of the lab after the wiki freeze
  • Sandra will ask for the appointment to talk at the MPI



green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Sandra, Sophie: primer design for NOT-gate to get NEH I restriction sites inside instead of Spe I. We can not cut with Spe because our trp-promoter contains Spe restriction sites.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

3A-assembly

1.Digestion

Investigators: Rüdiger


Sample name DNA concentration (ng/μl)
M61a 335
M61b 300
M62a 267
M62b 263
M63a 447
M63b 412

diegsted with EcoRI and PstI.


Gel extraction kit

Investigators: Theo

DNA fragments of the digestion( see above) were excised from the gel.

Ligation

Investigators: Rüdiger

Trying to ligate the precipitator (excised from the gel) in the pSB1C3 vector. Ratio of ligation: 1:3

                       

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