| Autor: |
Rahul Bhardwaj, Phan T. Tue, Manish Biyani, Yuzuru Takamura |
| Jazyk: |
angličtina |
| Rok vydání: |
2019 |
| Předmět: |
|
| Zdroj: |
Applied Sciences, Vol 9, Iss 7, p 1357 (2019) |
| Druh dokumentu: |
article |
| ISSN: |
2076-3417 |
| DOI: |
10.3390/app9071357 |
| Popis: |
Reverse DNA synthesis (5′-3′) plays diverse functional roles in cellular biology, biotechnology, and nanotechnology. However, current microfluidic systems for synthesizing single-stranded DNAs at a laboratory scale are limited. In this work, we develop a simple and efficient polydimethylsiloxane- (PDMS-) based microfluidic system for the reverse chemical synthesis of short-chain oligonucleotides (in the 5′-3′ direction) under ambient conditions. The use of a microfluidics device and anhydrous conditions effectively surpass the problem of moisture sensitivity during oligonucleotide synthesis. With optimized microfluidic synthesis conditions, the system is able to synthesize up to 21 bases-long oligonucleotides in air atmosphere. The as-synthesized oligonucleotides, without further purification, are characterized using matrix-assisted laser desorption ionization–time of flight (MALDI-TOF/TOF) mass spectroscopy (MS) supported by the denatured polyacrylamide gel electrophoresis (PAGE) analysis. This developed system is highly promising for producing the desired sequence at the nanomolar scale on-chip and on-demand in the near future. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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