Abstract:
:Modular primers are strings of three contiguously annealed unligated oligonucleotides (modules) as short as 5- or 6-mers, selected from a presynthesized library. It was previously found that such strings can prime DNA sequencing reactions specifically, thus eliminating the need for the primer synthesis step in DNA sequencing by primer walking. It has remained largely a mystery why modular primers prime uniquely, while a single module, used alone in the same conditions, often shows alternative priming of comparable strength. In a puzzling way, the single module, even in a large excess over the template, no longer primes at the alternative sites, when modules with which it can form a contiguous string are also present. Here we describe experiments indicating that this phenomenon cannot be explained by cooperative annealing of the modules to the template. Instead, the mechanism seems to involve competition between different primers for the available polymerase. In this competition, the polymerase is preferentially engaged by longer primers, whether modular or conventional, at the expense of shorter primers, even though the latter can otherwise prime with similar or occasionally higher efficiency.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Beskin AD,Zevin-Sonkin D,Sobolev IA,Ulanovsky LEdoi
10.1093/nar/23.15.2881subject
Has Abstractpub_date
1995-08-11 00:00:00pages
2881-5issue
15eissn
0305-1048issn
1362-4962pii
5w0078journal_volume
23pub_type
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