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Nucleic Acids Research, 1990, Vol. 18, No. 10 2891-2899
© 1990

MOLECULAR BIOLOGY

Cis-acting elements required for RNA polymerase II and III transcription in the human U2 and U6 snRNA promoters

Susan M. Lobo, Samantha Ifill and Nouria Hernandez

Cold Spring Harbor Laboratory Cold Spring Harbor, NY 11724, USA

Received February 14, 1990. Accepted April 9, 1990.

The recent development of yeast artificial chromosome (YAC) vectors has provided a system for cloning fragments that are over ten times larger than those that can be cloned in more established systems. We have developed a method for the rapid isolation of terminal sequences from YAC clones. The YAC clone is digested with a range of restriction enzymes, a common linker is ligated to the DNA fragments and terminal sequences are amplified using a vector specific primer and a linker specific primer. Sequence data derived from these terminal specific products can be used to design primers for a further round of screening to isolate overlapping clones. The method also provides a convenient method of generating Sequence Tagged Sites for the mapping of complex genomes.


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