Matt:LabNotes/2015-6-19

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Revision as of 21:45, 19 June 2015 by >Mzcai (Created page with "===DARTFISH with suppv2=== *Sample: 37C 10min **Labeled "DARTFISH 6.19.2015" #Wash the cells with 1ml of PBS (RNase free) 3 times #Prepare RT mixture '''on ice''' #*DEPC-H2O ...")
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DARTFISH with suppv2

  • Sample: 37C 10min
    • Labeled "DARTFISH 6.19.2015"
  1. Wash the cells with 1ml of PBS (RNase free) 3 times
  2. Prepare RT mixture on ice
    • DEPC-H2O - 157ul
    • M-MuLV RT Buffer 10X - 20ul
    • dNTP 25mM - 2ul
    • aa-dUTP 2mM - 4ul
    • RT primer 100uM - 5ul
      • Nonamer
    • RNase inhibitor 40U/ul - 2ul
    • M-MuLV reverse transcriptase 100U/ul - 10ul
  3. Add RT mixture and incubate for 10min at 4C
  4. Transfer sample to 37C overnight (~15hrs)