Simultaneous in situ detection of two mRNAs in the same cell using riboprobes labeled with biotin and 35S.
We used 35S-labeled and biotinylated cRNAs (riboprobes) to detect simultaneously two different mRNAs by in situ hybridization. In a first step we established the conditions under which each type of probe achieved the same high level of sensitivity. We then used these conditions to hybridize BHK cells infected with Theiler's virus, a murine picornavirus, with a mixture of a virus-specific biotinylated riboprobe and a 35S-labeled riboprobe specific for beta-actin mRNA. Both mRNAs could be detected in the same cell, although the sensitivity achieved by the radiolabeled probe was reduced by about 40% by the simultaneous hybridization with the biotinylated probe.
Unité Virus Lents, Institut Pasteur, Paris, France.
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S
Ozden
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Aubert
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Gonzalez-Dunia
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Brahic
D000199:Actins / Q000235:genetics; D000469:Alkaline Phosphatase; D000818:Animals; D001426:Bacterial Proteins; D001710:Biotin; D002460:Cell Line; D006224:Cricetinae; D007668:Kidney / Q000032:analysis / Q000166:cytology* / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D010849:Picornaviridae / Q000235:genetics; D010850:Picornaviridae Infections / Q000235:genetics; D012313:RNA / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D019809:Streptavidin; D013462:Sulfur Radioisotopes
D000199:Actins; D001426:Bacterial Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D013462:Sulfur Radioisotopes; D012313:RNA; D001710:Biotin; D019809:Streptavidin; D000469:Alkaline Phosphatase