Blue:RNA-Seq Experiments:Cordycepin test
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RNA-Seq Library Information[edit]
Samples | Condition |
TSO.r05 | 0/1 Cordycepin/ATP Enzymatics PAP |
TSO.r05 | 0.01/1 Cordycepin/ATP Enzymatics PAP |
TSO.r05 | 0.05/1 Cordycepin/ATP Enzymatics PAP |
TSO.r05 | 0.1/1 Cordycepin/ATP Enzymatics PAP |
TSO.r05 | 0.5/1 Cordycepin/ATP Enzymatics PAP |
TSO.r05 | 1/1 Cordycepin/ATP Enzymatics PAP |
TSO.r05 | 0/1 Cordycepin/ATP Affymetrix PAP |
TSO.r05 | 0/0 Cordycepin/ATP Enzymatics PAP |
Purpose[edit]
- Test Enzymatics versus Affymetrixs PAP
- Test Blocking of PAP activity using Cordycepin-ATP
Protocol[edit]
- Cordycepin Preparation
Mix equal volumes 2mM ATP with:
- Water = 0/1
- 0.02mM Cordycepin = 0.01/1
- 0.1mM Cordycepin = 0.05/1
- 0.2mM Cordycepin = 0.1/1
- 1mM Cordycepin = 0.5/1
- 2mM Cordycepin = 1/1
- 3' Tailing
Volume 1uM TSO.r05 1ul 5x RT Buffer 0.5ul cordycepin/ATP 0.25ul 0.5U/ul PAP (1:10d in H2O) 0.25ul H2O 2ul
- Incubate @ 37C 5 min
- Transfer to Ice
- Run 5ul on urea gel: