Six base oligonucleotide linkers and methods for their use
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United States of America Patent
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Jan 12, 1988
Grant Date -
N/A
app pub date -
Nov 30, 1984
filing date -
Nov 30, 1984
priority date (Note) -
Expired
status (Latency Note)
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Abstract
A method for inserting a selected restriction site into a double stranded genetic sequence at a selected tab site, and a linker for use therewith are disclosed. The method involves treating the genetic sequence with an opening agent (such as a restriction enzyme) so as to open both strands of the tab site. One then exposes the opened genetic sequence to two hexameric single stranded oligonucleotide linkers in the presence of a ligating enzyme. The two linkers have the same nitrogenous base sequence, a sequence which is partially palindromic complementary to itself at least one end of the nitrogenous sequence, but is not completely palindromic complementary to itself. Through use of this method, the two linkers are inserted in the opening with partial complementary overlap, and at least one of the linkers is affixed to each strand. Subsequently, the site is reclosed at the point of the insertion so as to contain a six base insertion.
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