| Title | Termination of DNA synthesis by novel 3'-modified-deoxyribonucleoside 5'-triphosphates. |
| Publication Type | Journal Article |
| Year of Publication | 1994 |
| Authors | Metzker, ML, Raghavachari, R, Richards, S, Jacutin, SE, Civitello, A, Burgess, K, Gibbs, RA |
| Journal | Nucleic Acids Res |
| Volume | 22 |
| Issue | 20 |
| Pagination | 4259-67 |
| Date Published | 1994 Oct 11 |
| ISSN | 0305-1048 |
| Keywords | Base Sequence, Chromatography, High Pressure Liquid, Deoxyribonucleotides, DNA, DNA-Directed DNA Polymerase, Molecular Sequence Data, Molecular Structure, Templates, Genetic |
| Abstract | Eight 3'-modified-dNTPs were synthesized and tested in two different DNA template assays for incorporation activity. From this enzymatic screen, two 3'-O-methyl-dNTPs were shown to terminate DNA syntheses mediated by a number of polymerases and may be used as alternative terminators in Sanger sequencing. 3'-O-(2-Nitrobenzyl)-dATP is a UV sensitive nucleotide and was shown to be incorporated by several thermostable DNA polymerases. Base specific termination and efficient photolytic removal of the 3'-protecting group was demonstrated. Following deprotection, DNA synthesis was reinitiated by the incorporation of natural nucleotides into DNA. The identification of this labile terminator and the demonstration of a one cycle stop-start DNA synthesis are initial steps in the development of a novel sequencing strategy.
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| DOI | 10.1093/nar/22.20.4259 |
| Alternate Journal | Nucleic Acids Res |
| PubMed ID | 7937154 |
| PubMed Central ID | PMC331938 |
| Grant List | 5 P30 HG00210 / HG / NHGRI NIH HHS / United States |