Incorporation of nonbase residues into synthetic oligonucleotides and their use in the PCR.
Oligonucleotides containing the nonbase residues 1,3-propanediol or 1,4-anhydro-2-deoxy-D-ribitol were synthesized and used as primers for the polymerase chain reaction (PCR). Since these residues cannot be replicated by a DNA polymerase, the resulting PCR products have protruding 5' ends. Primers were designed with three regions, a 3' region complementary to the desired template, a 5' region complementary to a preselected nucleotide sequence, and a nonreplicable element interposed between these two containing 1-3 of the nonbase residues. The primers were used in a PCR and the products hybridized without denaturation to a solid support containing an immobilized preselected nucleotide sequence. Studies are reported showing the effects of the nonreplicable elements in primer extension reactions and the application to the capture of PCR products.
Gade
R
R
Department of Molecular Biochemistry, Beckman Research Institute of the City of Hope, Duarte, California.
Kaplan
B E
BE
Swiderski
P M
PM
Wallace
R B
RB
D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism; D003855:Deoxyribose / Q000031:analogs & derivatives; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000138:chemical synthesis*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011409:Propylene Glycols
D017931:DNA Primers; D009841:Oligonucleotides; D011409:Propylene Glycols; D003855:Deoxyribose; C041787:1,3-propanediol; D004247:DNA; C043138:1,2-dideoxyribofuranose; D004259:DNA-Directed DNA Polymerase