Blue:RNA-Seq Experiments:Cordycepin test: Difference between revisions

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{| {{table}}
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Samples'''
| align="center" style="background:#f0f0f0;"|'''Samples'''
| align="center" style="background:#f0f0f0;"|'''Input RNA'''
| align="center" style="background:#f0f0f0;"|'''Condition'''
|-
|-
| TSO.r05||0/1 Cordycepin/ATP Enzymatics PAP
| TSO.r05||0/1 Cordycepin/ATP Enzymatics PAP

Revision as of 23:37, 18 October 2013

RNA-Seq Library Information

Samples Condition
TSO.r05 0/1 Cordycepin/ATP Enzymatics PAP
TSO.r05 0/1 Cordycepin/ATP Affymetrix 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

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 urea gel: