Matt:JustinLabNotes/2015-10-7

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Testing "Homemade" Reagents to be used with CircLigase II[edit]

  • CircLigase II kit does not provide enough 5M Betaine, 50mM MnCl2, and 10X CircLigaseII Buffer for DARTFISH volumes
  • Test 10X Buffer Hosuk made
  • Test 5M Betaine Justin made from dry (seems very old)
  • Test 50mM MnCl2 Justin made from Manganese chloride tetrahydrate (also seems very old from Paola's lab)

Protocol[edit]

  • Followed protocol provided by CircLigase II kit and control ssDNA in kit
    • Try substituting one "homemade" reagent for kit provided reagent in each sample

Results[edit]

File:20151007 CircLigaseIIReagentsTest.jpg

  • Hosuk's buffer works well
  • 5M Betaine by Justin does not work
    • Use 5M Betaine from SigmaAldrich
  • 50mM MnCl2 works decently