A rapid and convenient method to prepare DIG-labelled RNA probes for use in non-radioactive in situhybridization

We describe here the use of PCR-generated templates incorporating T3 polymerase sites in order to prepare digoxigenin (DIG)-labelled cRNA probes against any gene of known sequence. This method was applied to the preparation of probes specific for chicken glyceraldehyde-3-phosphate dehydrogenase mess...

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Bibliographic Details
Published inMolecular and cellular probes Vol. 10; no. 1; pp. 51 - 55
Main Authors Gandrillon, Olivier, Solari, Florence, Legrand, Claude, Jurdic, Pierre, Samarut, Jacques
Format Journal Article
LanguageEnglish
Published Elsevier Ltd 1996
Subjects
cRNA probe
digoxigenin
PCR
digoxigenin
PCR
cRNA probe
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Summary:We describe here the use of PCR-generated templates incorporating T3 polymerase sites in order to prepare digoxigenin (DIG)-labelled cRNA probes against any gene of known sequence. This method was applied to the preparation of probes specific for chicken glyceraldehyde-3-phosphate dehydrogenase messenger RNAs and we demonstrate that such probes can be used for in situhybridization (ISH). This technique therefore represents a rapid and convenient means to prepare DIG-labelled cRNA probes for use in a non-radioactive ISH. It adds speed and convenience of probe preparation to the previously described advantages of non-radioactive detection techniques.
ISSN:0890-8508
1096-1194
DOI:10.1006/mcpr.1996.0007