Team:TorontoMaRSDiscovery/11 June 2009


June 11, 2009

  • We did not have enough isolated plasmid DNA to justify doing another gel so we decided to perform another gel so we decided to perform another miniprep to get some more
  • Used DB3.1 transformed bacteria from shaker(measured spec 0.794)
  • Miniprep
    • potential issue our microcentrifuge only spins @ 10,000 not required 12,000
  • Binding DNA
    • did both optional steps preheated TE + washed w/ W10
  • measured UV absorbance = 4.2ng/ul
  • in all there is ~ 74 ul of TE should indicated ~370 ng of plasmid.
  • Digest
    • NOTE: plasmid length w/e biobric is 3189 bp biobrick length is 3255bp this can explain lack of 2 clear bands on gel after ligation.
    • nevertheless, we will perform another Gel to make sure we can properly run ladder to determine fragment sizes
  • digest measurements: 250ng plasmid all else half except BSA