Isolation of exons from cloned DNA by exon trapping.
Exon trapping is an RNA polymerase chain reaction (PCR) method to clone expressed sequences or exons directly from mammalian genomic DNA. The basic protocol in this unit describes the method for trapping internal exons from cosmid clones and the second basic protocol describes trapping of 3 terminal exons. An describes 3 terminal exon trapping, which avoids subcloning of target DNA by ligating it to the vector for direct transfection. A describes a rapid cloning procedure using uracil DNA glycosylase.
Nisson
P E
PE
Life Technologies, Gaithersburg, Maryland, USA.
Watkins
P C
PC
Krizman
D B
DB
D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D003360:Cosmids; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005091:Exons; D015723:Gene Library; D005821:Genetic Techniques; D005822:Genetic Vectors; D005826:Genetics, Medical; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*
D018076:DNA, Complementary