Reference - Detail
RRC ID | 35202 |
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Author | Celesnik H, Deana A, Belasco JG. |
Title | PABLO analysis of RNA: 5'-phosphorylation state and 5'-end mapping. |
Journal | Methods Enzymol |
Abstract |
Recent studies have revealed that 5'-end-dependent RNA degradation in prokaryotes is triggered by pyrophosphate removal from the 5'-terminus to generate a monophosphorylated intermediate that is readily degraded. This chapter describes how to examine the 5'-phosphorylation state of any specific bacterial RNA by PABLO analysis. The method is based on the ability of monophosphorylated, but not triphosphorylated, RNA 5'-ends to undergo splinted ligation to a DNA oligonucleotide when juxtaposed by base pairing to a bridging oligonucleotide. PABLO analysis not only makes it possible to quantify the proportion of a particular RNA that is monophosphorylated in bacterial cells but also provides a more reliable method than primer extension for high-resolution mapping of RNA 5'-termini. |
Volume | 447 |
Pages | 83-98 |
Published | 2008-1-1 |
DOI | 10.1016/S0076-6879(08)02205-2 |
PII | S0076-6879(08)02205-2 |
PMID | 19161839 |
MeSH | Base Sequence Blotting, Northern Diphosphates / isolation & purification Electrophoresis, Polyacrylamide Gel Molecular Sequence Data Phosphorylation RNA, Messenger / chemistry RNA, Messenger / metabolism* Reverse Transcriptase Polymerase Chain Reaction |
IF | 1.394 |
Times Cited | 23 |
WOS Category | BIOCHEMICAL RESEARCH METHODS BIOCHEMISTRY & MOLECULAR BIOLOGY |
Resource | |
Prokaryotes E. coli | Review? |