A large-scale in situ hybridization system using an equalized cDNA library.
We have developed a large-scale in situ hybridization system in which all the procedures are carried out on a 96-well format: digoxigenin-labeled probes were synthesized from PCR-amplified templates, sections were mounted on 96-well plates, and hybridization and immunohistochemistry protocols were carried out in each well of the plates. This system in combination with equalized (normalized) cDNA library as the source for the probes enables us to identify the cellular distribution of many mRNAs in various tissues rapidly and efficiently. Thus, this system may be a novel cloning method of identify genes differentially expressed in many tissues. In addition, this system has a potential to be automated.
Komiya
T
T
ERATO, Japan Science and Technology Corporation (JST), 5-9-4 Tokodai, Tsukuba, 300-26, Japan. tkom@hccp.jst.go.jp
Tanigawa
Y
Y
Hirohashi
S
S
D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics* / Q000378:metabolism; D004076:Digoxigenin / Q000276:immunology / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D007150:Immunohistochemistry; D017403:In Situ Hybridization / Q000295:instrumentation / Q000379:methods*; D007421:Intestine, Small / Q000737:chemistry; D051379:Mice; D009693:Nucleic Acid Hybridization / Q000379:methods; D011506:Proteins / Q000032:analysis; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D017931:DNA Primers; D018076:DNA, Complementary; D011506:Proteins; D012333:RNA, Messenger; D004076:Digoxigenin