Team:TUDelft/16 July 2009

=16th July=

Sriram & Tim Vos
Got the error why kanamycin plates didn't show colonies. It seems I added twice the amount of kanamycin in plates. Since the stock antibiotics were of 1000x concentration, just diluting to 1x would be sufficient. For 20 ml plates just adding 20 &micro;l of any antibiotic would be sufficient. Thus all the kanamycin containing biobricks must be redone.

I got colonies in all the tubes except the GFP(E0040). So the colony scooped from the plate was not proper. So must be re-cultured. And stupid me, I labeled TetR (C0040) and cl (C0051) containing culture tubes as TetR. So I need to re-culture them from plates and oomph plates are getting overgrown colonies so was transferred fridge after covering with parafilm.

Me and Vos isolated the DNA from 8 of the cultures grown for biobricks [R0010, R0040, J23008, J23031, E1010, B0034, B0015, K081013] using MiniPrep Plasmid extraction kit in 3 hours and kept in -20&deg;C freezer.

At 5:30 PM I recultured the GFP(E0040), TetR(C0040) and cl(C0051) from their plates in 3 new culture tubes with 5&micro;l Ampicillin and kept in shaker at 160 rpm and 37 &deg;C.

Weenink
Purified I-SceI homing endonuclease arrived today in a box of dry ice. Nice! All my transformants gave colonies. The B0032 plate had lots of colonies with lots of sattelite colonies as well (not good). So I have started 5ml LB 1 x amp liquid cultures at 37ºC of B0032, K145015 and K145201. (inoculation at 18:20) Plates are now in the fridge. Also I backdiluted the K145280HS strain in 30 ml liquid culture. (same conditions as before, inoc at 18:15). Finally I plated out the ccdB resistant strains from last year (out of -80 stock) on LB 1xamp plates at 18:30. Plasmid names: pSB1AC3,pSB1AT3,pSB1AK3. incubated in 37ºC stove. To be grown up in LC and purified tomorrow.

Calin
Wrote a simplified Matlab script for the riboregulator scheme 1 which treats the secondary structure interactions as normal enzyme substrate interaction. Did some stability analysis. Added code to automatically calculate the delay time based on a predefined threshold. Drew the proposed team logo in vector format.