IGEM:Indian Institue of Technology Madras/2009/Notebook/PLASMID - Plasmid Locking Assembly for Sustaining Multiple Insert DNA/2009/09/01

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  • Following parts were digested again with EcoR1 (because only some of the parts digested on 31.8.09, namely ________ were digested properly):
  • J23119+S03879 (from 10 colonies)
  • K145151+B0014 (from 10 colonies)
  • J23119+E0420 (from 10 colonies)
  • R0010 +E0420 (from 4 colonies)
  • J13002 (from 5 colonies)

Note: In a reaction volume of 50 micro l, 10 micro l of intact DNA was used for digestion instead of the usual 5 micro l.

  • Following parts were mini prepped and digested with EcoR1:
  • R0010+E0420 (from 5 colonies)
  • K117005+K145151 (from 10 colonies)
  • All the digested parts and their intact counterparts were run on a gel.
  • Total wells loaded, excluding the ladders: (39 + 15)x2 = 108
  • 5 micro l of intact DNA and 10 micro l of digest for each part were loaded.


  • Order of loading the wells:
  • Gel_1, row 1: J23119+S03879(intact plasmids-from colonies 1 to 10 into wells 1 to 10)-ladder- J23119+S03879(digested by EcoR1-from colonies 1 to 10 into wells 12 to 21)
  • Gel_1, row 2: K145151+B0014(intact plasmids-from colonies 1 to 10 into wells 1 to 10)-ladder- K145151+B0014(digested by EcoR1-from colonies 1 to 10 into wells 12 to 21)
  • Gel_2, row 1: J23119+E0420(intact plasmids-from colonies 1 to 10 into wells 1 to 10)-ladder- J23119+E0420(digested by EcoR1-from colonies 1 to 10 into wells 12 to 21)
  • Gel_2, row 2: J13002 (intact plasmids- from colonies 1 to 5 into wells 1 to 5),R0010+E0420(intact plasmids- from colonies 1 to 4 into wells 6 to 9)-ladder- J13002 (digested with EcoR1- from colonies 1 to 5 into wells 11 to 15),R0010+E0420(digested with EcoR1- from colonies 1 to 4 into wells 16 to 19)
  • Gel_3, row 1:



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