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From 2012.igem.org

week 4: 21 may - 25 may

office work

This week feeled a bit of a slow week. Two of our team members were a day sick. After the great succes with the phyre2 program, we searched for a program that uses the output of the phyre2 program to predict the quaternary structure of the VLPs. In other words can a VLP be formed. We found out that there aren't many programs that can accuratly predict the quaternary structure and the programs that are there aren't really compatible with phyre2. Besides getting stuck with the prediction model Mark investigates further for a new method to determain if VLPs were formed. He found two methods that can be used: assymetric flow field-flow fractionation (AFFFF) followed by multi angle light scaterring (MALS) and dynamic light scattering (DLS). Thijs and Jasper continued further with the development of the deconstructor and are hoping that after the weekend the program is ready for beta testing.

[meeting]

written by: Mark

lab work

Testing CCMV protocol

Monday:

Sonified 6 x 30 seconds, output 3-4, amplitude around 20 on ice! cool as much possible, 50 seconds on ice between 30 second steps. lysate in 50 ml disassembly buffer.

Centrifuged at 15000 RPM, 3 minute 25 ml is dialised, 1st time, overnight.

Tuesday:

Negative control for SDS Non induced BL21 CCMV grew overnight in LB+kan follow protocol CCMV from day 1, after induction up to the 4th step of day 2

sample transferred to new tube stored in iGEM -20 degree Celcius as BL21 negative - iGEM SDS

Thursday:

BL21 Plated and greiner tube 10 ml tube

BL21 2 plates freezer BL21 2 Greiner tubes 30 degree Celcius stove