IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/15

From OpenWetWare
Jump to navigationJump to search
Main project page
Previous entry      

Check big stock of RBS S and W

  • We want to check our big stock of RBS S and W, with all digest enzymes

- Plasmid miniprep RBS S6 and RBS W6 (from 200mL flask)

- Nanodrop

product concentration (ng/μL) 260/280
RBS W2 90.3 1.70
RBS W4 124.0 1.80

- Single digest with EcoRI, XbaI, SpeI and PstI

- Gel

  • Lane1 : HyperladderI
  • Lane2 : RBS S6 cut with EcoRI
  • Lane3 : RBS S6 cut with SpeI
  • Lane4 : RBS S6 cut with PstI
  • Lane5 : RBS S6 cut with XbaI
  • Lane6 : RBS S6 uncut
  • Lane7 : RBS W6 cut with EcoRI
  • Lane8 : RBS W6 cut with SpeI
  • Lane9 : RBS W6 cut with PstI
  • Lane10 : RBS W6 cut with XbaI
  • Lane11 : RBS W6 uncut
  • Lane12 : supercoiled ladder
  • Result

- Even no band for the uncut plasmid. There is a problem with plasmid miniprep, we will try again with less (more diluted) cells for plasmid miniprep.

- New plasmid miniprep (the pellets from 20mL of LB is diluted into 1.2ml instead of 0.6)

- New single digest

- New gel (same lanes than before)

  • Result

- One band for everything, but the same size than the uncut plasmid. Moreover, this uncut plasmid should be 3000bp, and here, it is more. So, we have a problem in the growth of cells, we will try again and check the 10ml tube which is used to inoculate the big flask and also the big flask.


Retrieved from "https://openwetware.org/mediawiki/index.php?title=IGEM:Cambridge/2008/Notebook/Turing_Pattern_Formation/2008/09/15&oldid=1000042"