A sensitive dot blot procedure for detecting mRNA in lymphoid cells grown in liquid culture.
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.
Benveniste
P
P
Ontario Cancer Institute, Toronto, Canada.
Derry
H
H
Wu
G E
GE
Paige
C J
CJ
Miller
R G
RG
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
D012333:RNA, Messenger; D011948:Receptors, Antigen, T-Cell; D004247:DNA