Matt:LabNotes/2013-3-9

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Second Strand cDNA synthesis[edit]

  • Follow NEB E6111S protocol: [1]

Started with 20 ul cDNA from Friday 3/8/13

  • Put in ice
  • Added 48 ul H20
  • Added 8ul 10x Second strand synthesis reaction buffer
  • Added 4ul Second strand synthesis enzyme mix
  • Mixed by pipetting
  • Incubated for 2.5 hours at 16C
  • Purified using Qiagen minelute column (eluted 23ul into 1.5ml tube labelled HBRR cDNA)
  • Measured DNA conc with Nandrop: 55.5 ng/ul
    • 21.5ul (1.19ug) sample left in tube