Imperial/Wet Lab/Lab Notebook/2007-09-14

From 2007.igem.org

14 September 2007

Restriction Digest of Biobricks

  1. 4 DNA samples were digested by SpeI and PstI
    • Samples 5, 7, 24
  2. 1 DNA sample wasdigested by XbaI and PstI
    • Samples 23
  3. 4 DNA samples were digested by EcoRI
    • Samples 13, 25, 27b-e, T7, Kirsten's parts
  4. 4 μl of DNA was digested by 1 μl of enzymes at 37°C for 1 hour
    Exception: Sample 23 had 6 μl DNA and 2 μl enzymes

Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Restriction_Digest | Restriction Digest] in the general protocols page

Agarose Gel Electrophoresis of Biobricks

  1. Checked 9 Biobricks and 13 Digests on 1% agarose gel

ICGEMS Gel 14 9.png

Conclusions:

  • Part 27 is confirmed to be faulty
  • T7 maxi-prep was not done properly
  • Kirsten's parts were most likely not there...

Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Electrophoresis | Electrophoresis] in the general protocols page

Gel Purification of Biobricks

  1. Gel was melted and run through column
  2. Purified DNA was eluted with ddH2O

  • 5. BBa_R0040 [ptet] (~2100 bp fragment)
  • 7. BBa_F2620 [ptet-LuxR-pLux] (~3100 bp fragment)
  • 23. BBa_I13504 [GFP] (~800 bp fragment)
  • 24. BBa_R0062 [pLux] (~2100 bp fragment)

Protocols can be found at [http://www.openwetware.org/wiki/IGEM:IMPERIAL/2007/Notebook/General_Protocols#Gel_Purification | Gel Purification] in the general protocols page