Detection of CAIII mRNA in rat skeletal muscle and liver by in situ hybridization.
We carried out a variety of in situ methods of hybridization on rat liver and rat skeletal muscle using 35S-labeled or biotin-labeled rat carbonic anhydrase III (CAIII) cDNA clone. The methods were compared and evaluated. Use of the biotin system produced defined but nonspecific results which were shown not to be due to the biotinylated cDNA probe binding to the mRNA in the muscle sections. This artifact was shown to persist despite various attempts to eliminate it. Alternatively, using 35S-labeled cDNA gave reproducible results which were shown to be consistent with probe binding specifically to mRNA in the muscle section.
Kelly
C D
CD
Department of Child Health, St George's Hospital Medical School, London, United Kingdom.
Carter
N D
ND
de Boer
P
P
Jeffery
S
S
Moorman
A F
AF
Smith
A
A
D000251:Adenosine Triphosphatases / Q000032:analysis; D000818:Animals; D001710:Biotin; D002256:Carbonic Anhydrases / Q000032:analysis* / Q000235:genetics; D015342:DNA Probes; D008099:Liver / Q000201:enzymology*; D009132:Muscles / Q000201:enzymology*; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis*; D051381:Rats; D013462:Sulfur Radioisotopes
D015342:DNA Probes; D012333:RNA, Messenger; D013462:Sulfur Radioisotopes; D001710:Biotin; D000251:Adenosine Triphosphatases; D002256:Carbonic Anhydrases