Abstract:
:Oligonucleotides synthesized in array format suffer from contamination by truncated species. We have developed a method to invert DNA molecules in situ after completed synthesis. Reactive functions at the 5'-ends of the oligonucleotides are permitted to react with functions on the support before the 3'-ends are released, in effect reversing the orientation of full-length oligonucleotides, while any 5'-truncated molecules are lost. This strategy serves both to purify in situ synthesized reagents and to reorient the oligonucleotides, causing them to expose free 3'-hydroxyls. In situ inverted oligonucleotides can be used in assays based on DNA polymerase-assisted extension of immobilized primers, and we demonstrate their utility in minisequencing and in pyrosequencing.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Kwiatkowski M,Fredriksson S,Isaksson A,Nilsson M,Landegren Udoi
10.1093/nar/27.24.4710keywords:
subject
Has Abstractpub_date
1999-12-15 00:00:00pages
4710-4issue
24eissn
0305-1048issn
1362-4962pii
gkc680journal_volume
27pub_type
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