Blue:RNA-Seq Experiments:Cordycepin test

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RNA-Seq Library Information

Samples Input RNA TSO Primer
TSO.r05 0/1 Cordycepin/ATP Enzymatics PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read
TSO.r05 0/1 Cordycepin/ATP Affymetrix PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read
TSO.r05 0.01/1 Cordycepin/ATP Enzymatics PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read
TSO.r05 0.05/1 Cordycepin/ATP Enzymatics PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read
TSO.r05 0.1/1 Cordycepin/ATP Enzymatics PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read
TSO.r05 0.5/1 Cordycepin/ATP Enzymatics PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read
TSO.r05 1/1 Cordycepin/ATP Enzymatics PAP TSO.r06 T20V.id N2.id] totoRNAseq Read1 plus N2 barcode read


Purpose

  • Test Enzymatics versus Affymetrixs PAP
  • Test Blocking of PAP activity using Cordycepin-ATP


Protocol

Cordycepin Preparation

Mix equal volumes 2mM ATP with:

  1. Water = 0/1
  2. 0.02mM Cordycepin = 0.01/1
  3. 0.1mM Cordycepin = 0.05/1
  4. 0.2mM Cordycepin = 0.1/1
  5. 1mM Cordycepin = 0.5/1
  6. 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 gel: