Visualization and quantitation of poly A+-mRNA by using in situ hybridization with biotinylated oligonucleotides.
HeLa cells and synovial tissue sections were used for visualizing poly A+-mRNA after in situ hybridization. We describe a method to biotinylate hybridization probes by tailing oligo-dT nucleotides with biotinyl-dUTP using terminal deoxynucleotidyl transferase (E.C.2.7.7.3.1.). The hybrids were detected by using the strepavidin-biotin-peroxidase complex (ABC method). The reaction product of the peroxidase staining in the cells and in the tissue sections, is quantitated by the method of plug- and scanning photometry respectively.
Wehner
K
K
Biologisches Institut der Universität Stuttgart.
Fritz
P
P
Köhler
K
K
Multhaupt
H
H
D006367:HeLa Cells; D006801:Humans; D007124:Immunoenzyme Techniques; D009363:Neoplasm Proteins / Q000096:biosynthesis; D009693:Nucleic Acid Hybridization; D015345:Oligonucleotide Probes; D011061:Poly A / Q000032:analysis*; D012333:RNA, Messenger / Q000032:analysis*
D009363:Neoplasm Proteins; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011061:Poly A