title: A sensitive dot blot procedure for detecting mRNA in lymphoid cells grown in liquid culture.
abstract: A sensitive dot blot procedure for detecting RNA transcripts with radiolabelled cDNA probes in small numbers of cells in suspension culture is described. For a frequent mRNA species such as that for immunoglobulin in an IgM secretor hybridoma, as few as 30 cells can be detected using a C mu probe; for a less frequent mRNA species such as transcripts coding for alpha, beta or gamma genes of the T cell receptor in T cell tumors or CTL clones, as few as 3 X 10(3) to 10(4) cells can be detected using C alpha, C beta or C gamma probes. High sensitivity is achieved by confining the cells to a very small area on the filter on which they are probed, and by treating the filter with formaldehyde.
authors.affiliation: Ontario Cancer Institute, Toronto, Canada.
authors.forename: P
authors.initials: P
authors.lastname: Benveniste
authors.forename: H
authors.initials: H
authors.lastname: Derry
authors.forename: G E
authors.initials: GE
authors.lastname: Wu
authors.forename: C J
authors.initials: CJ
authors.lastname: Paige
authors.forename: R G
authors.initials: RG
authors.lastname: Miller
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D004247:DNA / Q000235:genetics; D005796:Genes; D005803:Genes, Immunoglobulin; D008214:Lymphocytes / Q000276:immunology*; D051379:Mice; D008815:Mice, Inbred Strains; D008819:Mice, Nude; D009693:Nucleic Acid Hybridization; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology; D014158:Transcription, Genetic
chemical_list: D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D004247:DNA