Accurate and Efficient One-Pot Reverse Transcription and Amplification of 2′-Fluoro-Modified Nucleic Acids by Commercial DNA Polymerases
| Autor: | Aurora G. Weiden, Madison K. C. Seto, Ananya Venkatesh, Simone Z. P. Gottlieb, Aaron M. Leconte, Arianna S. Thompson, Susanna E. Barrett |
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| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular chemistry.chemical_classification Halogenation biology Chemistry DNA polymerase fungi food and beverages DNA DNA-Directed DNA Polymerase Reverse Transcription Biochemistry Reverse transcriptase Synthetic biology chemistry.chemical_compound Enzyme Nucleic acid biology.protein Nucleic Acid Conformation Nucleic Acid Amplification Techniques |
| Zdroj: | Biochemistry. 59:2833-2841 |
| ISSN: | 1520-4995 0006-2960 |
| Popis: | DNA is a foundational tool in biotechnology and synthetic biology but is limited by sensitivity to DNA-modifying enzymes. Recently, researchers have identified DNA polymerases that can enzymatically synthesize long oligonucleotides of modified DNA (M-DNA) that are resistant to DNA-modifying enzymes. Most applications require M-DNA to be reverse transcribed, typically using a RNA reverse transcriptase, back into natural DNA for sequence analysis or further manipulation. Here, we tested commercially available DNA-dependent DNA polymerases for their ability to reverse transcribe and amplify M-DNA in a one-pot reaction. Three of the six polymerases chosen (Phusion, Q5, and Deep Vent) could reverse transcribe and amplify synthetic 2'F M-DNA in a single reaction with5 × 10 |
| Databáze: | OpenAIRE |
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