Multiplex detection of RNA expression in Drosophila embryos.
We present a fluorescence-based, multiplex in situ hybridization method that permits the simultaneous detection of five differently labeled antisense RNA probes and up to seven differ-ent transcripts in a single Drosophila embryo. We also show that it should be possible to increase the number of detected transcripts substantially with nascent transcript multiplex fluorescent in situ hybridization. These multiplex methods fill a current technological gap between high-resolution in situ hybridization with one or two fluorescently labeled probes and low-resolution but genome-wide microarray RNA profiling and should be of great utility in establishing gene networks.
Kosman
Dave
D
Section of Cell and Developmental Biology, University of California-San Diego, La Jolla, CA 92093, USA.
Mizutani
Claudia M
CM
Lemons
Derek
D
Cox
W Gregory
WG
McGinnis
William
W
Bier
Ethan
E
D000818:Animals; D004330:Drosophila / Q000196:embryology / Q000235:genetics*; D004625:Embryo, Nonmammalian / Q000502:physiology*; D005456:Fluorescent Dyes; D015870:Gene Expression; D020869:Gene Expression Profiling / Q000379:methods*; D017344:Genes, Insect; D017404:In Situ Hybridization, Fluorescence; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D014158:Transcription, Genetic
D005456:Fluorescent Dyes; D015347:RNA Probes; D012333:RNA, Messenger