Increasing proportions of uracil in DNA substrates increases inhibition of restriction enzyme digests
Techniques that rely upon the incorporation of uracil into DNA are being published with increasing frequency, especially in PCR protocols. We report here the efficiency of 18 type II restriction enzymes to digest PCR amplicons synthesized with varying proportions of TTP to dUTP in the PCR mixture. W...
| Publié dans: | BioTechniques. - 1991. - 17(1994), 6 vom: 15. Dez., Seite 1086-90 |
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| Auteur principal: | |
| Autres auteurs: | , |
| Format: | Article |
| Langue: | English |
| Publié: |
1994
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| Accès à la collection: | BioTechniques |
| Sujets: | Technical Report Journal Article Thymine Nucleotides Uracil Nucleotides DNA Restriction Enzymes EC 3.1.21.- |
| Résumé: | Techniques that rely upon the incorporation of uracil into DNA are being published with increasing frequency, especially in PCR protocols. We report here the efficiency of 18 type II restriction enzymes to digest PCR amplicons synthesized with varying proportions of TTP to dUTP in the PCR mixture. We find that most enzymes with A:T/U bp in their recognition site digest the amplicons less efficiently as the percentage of dUTP in the reaction mixture is increased. This effect is most dramatic when the proportion of dUTP in the nucleotide mixture exceeds 50%. All but one of the enzymes which fail to digest amplicons that are synthesized with 100% dUTP digest some amplicons which are synthesized with 90% dUTP |
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| Description: | Date Completed 31.03.1995 Date Revised 15.11.2006 published: Print Citation Status MEDLINE |
| ISSN: | 1940-9818 |