Amplification and Direct Sequence Analysis of the 23S rRNA Gene from Thermophilic Bacteria
| Autor: | Ashraf Ibrahim, Jacob Hofman-Bang, Birgitte K. Ahring |
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| Jazyk: | angličtina |
| Rok vydání: | 2001 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 30, Iss 2, Pp 414-420 (2001) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/01302rr05 |
| Popis: | We present a simplified and fast method to obtain high-quality sequences directly from PCRs without the traditional gel purification. We also report on an improved method to obtain sequence-quality PCR products from microorganisms that are difficult to lyse with no need for DNA extraction. The technique uses exonuclease I and shrimp alkaline phosphatase to degrade residual dNTPs and primers. Our technique is shown to work on both Gram-positive and Gram-negative bacteria. |
| Databáze: | Directory of Open Access Journals |
| Externí odkaz: |
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