High-throughput production of optimized primers (fimers) for whole-genome direct sequencing.
Fimers are specifically modified primers selected to inhibit nonspecific interactions occurring in cycle sequencing. They are postsynthetically derived from 2'-methoxyoxalamido or 2'-succinimido precursor oligonucleotides by treatment with appropriate small molecular weight modifiers (a primary aliphatic amine or hydroxide anion). We describe design, synthesis, and purification of fimers, and their use in protocols for direct sequencing of genomic deoxyribonucleic acid (DNA). Protocol for isolation of microbial genomic DNA is also reported.
Polushin
Nikolai
N
Fidelity Systems Inc., Gaithresburg, MD, USA
Malykh
Andrei
A
Morocho
A Michael
AM
Slesarev
Alexei
A
Kozyavkin
Sergei
S
D001483:Base Sequence; D017931:DNA Primers; D016678:Genome
D017931:DNA Primers