Blue:RNA-Seq Experiments:10232013: Difference between revisions
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>B1lake (Created page with " == Purpose == *Test PAP Activity from different companies: **Ambion **Enzymatics (used up to this point) **Affymetrix *Test Blocking of PAP activity using Cordycepin-ATP =...") |
>B1lake |
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:::''Incubate @ 37C 0, 5, 10, 15, 20 min'' | :::''Incubate @ 37C 0, 5, 10, 15, 20 min'' | ||
:::''Transfer to Ice'' | :::''Transfer to Ice'' | ||
*Run 5ul on urea gel: | *Run 5ul on urea gel: | ||
[[File:10232013_gel1.jpg]] |
Revision as of 23:50, 23 October 2013
Purpose
- Test PAP Activity from different companies:
- Ambion
- Enzymatics (used up to this point)
- Affymetrix
- Test Blocking of PAP activity using Cordycepin-ATP
Protocol
- Ambion 3' Tailing
Volume 1uM TSO.r04 1ul dH20 4.6ul 5x E-PAP buffer 2ul 25mM MnCl2 1ul 10mM ATP 1ul 2U/ul PAP 0.4ul
- Enzymatics 3' Tailing
Volume 1uM TSO.r04 1ul dH20 6ul 10x PAP buffer 1ul 10mM ATP 1ul 0.5U/ul PAP (1:10 Dil) 1ul
- Affymetrix 3' Tailing
Volume 1uM TSO.r04 1ul dH20 5ul 5x PAP buffer 2ul 10mM ATP 1ul 60U/ul PAP (1:10 Dil) 1ul
- Incubate @ 37C 0, 5, 10, 15, 20 min
- Transfer to Ice
- Run 5ul on urea gel: