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  4. 3'-end labeling of RNA with recombinant yeast poly(A) polymerase
 
research article

3'-end labeling of RNA with recombinant yeast poly(A) polymerase

Lingner, J.  
•
Keller, W.
1993
Nucleic Acids Res

Two commonly used methods to end-label RNA-molecules are 5'-end labeling by polynucleotide kinase and 3'-end labeling with pCp and T4 RNA ligase. We show here that RNA 3'-ends can also be labeled with the chain-terminating analogue cordycepin 5'-triphosphate (3'-deoxy-ATP) which is added by poly(A) polymerase. For a synthetic RNA it is shown that 40% of cordycepin becomes incorporated when the nucleotide is used at limiting concentrations and that with an excess of cordycepin 5'-triphosphate essentially all the RNA becomes modified at its 3'-end. The reaction is complete within minutes and the RNA product is uniform and suitable for sequence analysis. The efficiency of labeling varies with different RNA-molecules and is different from RNA ligase. Poly(A) polymerase preferentially labels longer RNA-molecules whereas short RNA-molecules are labeled more efficiently by T4 RNA ligase.

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Type
research article
DOI
10.1093/nar/21.12.2917
Author(s)
Lingner, J.  
Keller, W.
Date Issued

1993

Published in
Nucleic Acids Res
Volume

21

Issue

12

Start page

2917

End page

20

Note

Department of Cell Biology, Biozentrum of the University of Basel, Switzerland.

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
UPLIN  
Available on Infoscience
November 20, 2007
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/14755
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