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of Oligonucleotides by ESI-MS I 559
| Content Provider | Scilit |
|---|---|
| Author | Brown, Phyllis R. |
| Copyright Year | 2000 |
| Description | Another problem arises when determining PCR products with mass spectrometry, namely that the commonly used DNA-polymerase enzyme Taq can transfer a nontemplated adenine nucleotide dA to the 3' end of the PCR products because of its terminal deoxynucleotidyl transferase activity [60]. This causes the PCR products to have a nonuniform molecular weight and complicates the interpretation of the MS data. A solution to this problem was found by incorporating an EcoRI recognition site in the PCR primers [45-47]. Upon digestion of the PCR products with EcoRI, products of uniform molecular weight were obtained. This is a complication in the analysis and an additional time loss. A better solution was found by Muddiman et al. [12]. They used the DNA polymerase from Pyrococcus furiosus (Pfu), which does not incorporate a nontemplated adenine nucleotide (Fig. 7). Using Pfu, no additional digestion was necessary. Book Name: Advances in Chromatography |
| Related Links | https://content.taylorfrancis.com/books/download?dac=C2010-0-31828-2&isbn=9780429180668&doi=10.1201/9781482289800-67&format=pdf |
| Ending Page | 588 |
| Page Count | 2 |
| Starting Page | 587 |
| DOI | 10.1201/9781482289800-67 |
| Language | English |
| Publisher | Informa UK Limited |
| Publisher Date | 2000-02-09 |
| Access Restriction | Open |
| Subject Keyword | Book Name: Advances in Chromatography Critical Care Medicine Pcr Products Nontemplated Adenine Nucleotide Used the Dna Polymerase |
| Content Type | Text |
| Resource Type | Chapter |
