Isolation of genes from complex sources of mammalian genomic DNA using exon amplification.
Modifications to exon amplification have been instituted that increase its speed, efficiency and reliability. Exons were isolated from target human or mouse genomic DNA sources ranging from 30 kilobases (kb) to 3 megabases (Mb) in complexity. The efficiency was dependent upon the amount of input DNA, and ranged from isolation of an exon for every 20 kb to an exon for every 80 kb of target genomic DNA. In these studies, several novel genes and a smaller number of genes isolated previously that reside on human chromosome 9 have been identified. These results indicate that exon amplification is presently adaptable to large scale isolation of exons from complex sources of genomic DNA.
Church
D M
DM
Molecular Neurogenetics Unit, Massachusetts General Hospital, Boston.
Stotler
C J
CJ
Rutter
J L
JL
Murrell
J R
JR
Trofatter
J A
JA
Buckler
A J
AJ
D000818:Animals; D001483:Base Sequence; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000235:genetics; D002899:Chromosomes, Human, Pair 9; D004247:DNA / Q000235:genetics* / Q000302:isolation & purification; D017931:DNA Primers / Q000235:genetics; D005091:Exons; D005784:Gene Amplification; D005821:Genetic Techniques; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D004247:DNA; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase