Lab Notebook

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 µl 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°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 5µl Ampicillin and kept in shaker at 160 rpm and 37 °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.