Team:HokkaidoU Japan/Notebook/August2

From 2010.igem.org

Restriction enzyme digestion

Added 30 uL of TE into the DNA solution (purified via miniprep)

Digestion Menu

sample 1 sample 2 sample 3 sample 4
DNA solution 1 uL 1 uL 1 uL 1 uL
10x H Buffer - 1 uL 1 uL 1 uL
DW 9 uL 7.5 uL 7.0 uL 7.5 uL
Xba1 - 0.5 uL 0.5 uL -
Pst1 - - 0.5 uL 0.5 uL


  1. incubated at 37C for 1 hrs
  2. add 2 uL of 6x sample buffer
  3. Electrophoresed

Result of Electrophoresis

HokkaidoU Japan 20100802b.jpg
  • The insert lenght wants 1069 bp (plus RE recognition site, 49 bp)
  • Plasmid backbone was 5263 bp
  • Marker was λ/Hind III and pUC119/Hinf I
Good