Hosuk:LabNotes/2014-5-2: Difference between revisions

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(Created page with "*LabNote ===Next Plan=== ====Goal==== *Increase the Yield of cDNA Rolonies – from mRNAs *Enhance the yield of cDNA and Target-Specific RT ====How?===...")
 
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*Test long annealing time for RT primer
*Test long annealing time for RT primer
**Long Cai hybridized his FISH probes overnight
**Long Cai hybridized his FISH probes overnight
*Example of Test condition : 16 samples
**+a-dUTP, -a-dUTP
**Primer : Target-Specific Primer, Random Hexamer
**RT Primer annealing time : 15min, 1hr., 6hr., 12hr.

Latest revision as of 17:36, 7 May 2014

Next Plan[edit]

Goal[edit]

  • Increase the Yield of cDNA Rolonies – from mRNAs
  • Enhance the yield of cDNA and Target-Specific RT


How?[edit]

  • Apply the latest version of Rolony protocol (Nat. Protocol paper)
    • There are a few of changes such as fixation and RT conditions, amount of enzymes
  • Test +/- a-dUTP to increase RT efficiency
  • Test long annealing time for RT primer
    • Long Cai hybridized his FISH probes overnight


  • Example of Test condition : 16 samples
    • +a-dUTP, -a-dUTP
    • Primer : Target-Specific Primer, Random Hexamer
    • RT Primer annealing time : 15min, 1hr., 6hr., 12hr.