Strategy for chromosomal assignment of expressed sequences derived from heteronuclear RNA.
The chromosomal localization of transcribed sequences/genes is one of the objectives of the human genome project. Here, we describe a novel strategy for fast and dependable chromosomal assignment of expressed sequences that contain Alu sequences. Alu-PCR was performed on cDNA that was derived from heteronuclear (hn) RNA. hn-cDNA libraries are utilized for the identification of genes from extended human chromosomal regions or entire chromosomes. For chromosomal assignment, Alu-PCR products larger than 500 bp were hybridized against genomic DNA of somatic cell hybrids that was also amplified by Alu-PCR. Hybridization signals obtained within 2-3 h of exposure allow localization of cDNA-derived Alu-PCR products to single chromosomes. This technique is particularly useful for the analysis of cDNA libraries derived from hn-RNA. Using hn-cDNA clones from different chromosomes, we demonstrate the accuracy and reliability of the mapping strategy.
Brass
N
N
University of Saarland, Homburg/Saar, FRG.
Fischer
U
U
Mueller
H W
HW
Klein
V
V
Meese
E
E
D000818:Animals; D002874:Chromosome Mapping / Q000379:methods*; D006224:Cricetinae; D018076:DNA, Complementary / Q000235:genetics; D006801:Humans; D006822:Hybrid Cells; D016133:Polymerase Chain Reaction; D012332:RNA, Heterogeneous Nuclear / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid
D018076:DNA, Complementary; D012332:RNA, Heterogeneous Nuclear