PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA
We formulated a method to synthesize 1 kbp DNA fragments using ‘oligomer unidirectional joining method’ via asymmetric extension supported by a simulator for oligonucleotide extension (AESOE). In this study, trials were conducted on 41 sets of different genomic pieces of ten flaviviral genomes, and...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Future Science Ltd
2023-06-01
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| Series: | BioTechniques |
| Subjects: | |
| Online Access: | https://www.future-science.com/doi/10.2144/btn-2022-0127 |
| _version_ | 1797772981169029120 |
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| author | Yasunori Nishida Kotetsu Kayama Taichi Endoh Kiwamu Hanazono Gerry Amor Camer Daiji Endoh |
| author_facet | Yasunori Nishida Kotetsu Kayama Taichi Endoh Kiwamu Hanazono Gerry Amor Camer Daiji Endoh |
| author_sort | Yasunori Nishida |
| collection | DOAJ |
| description | We formulated a method to synthesize 1 kbp DNA fragments using ‘oligomer unidirectional joining method’ via asymmetric extension supported by a simulator for oligonucleotide extension (AESOE). In this study, trials were conducted on 41 sets of different genomic pieces of ten flaviviral genomes, and 31 bacterial 16s rRNA fragments with sizes ranging from 500 bases to 1.0 kbp. Synthetic gene production was found to be successful in all those sets. The synthesis method has three steps: the first step is a seven-linked AESOE, the second step is the linking of the 400-base fragments from the first step, and the third step is the final amplification. Our present approach is highly reproducible and may no longer require optimization of oligomer design. |
| first_indexed | 2024-03-12T21:59:35Z |
| format | Article |
| id | doaj.art-fbf481aee640489db6e1383d79b060a2 |
| institution | Directory Open Access Journal |
| issn | 0736-6205 1940-9818 |
| language | English |
| last_indexed | 2024-03-12T21:59:35Z |
| publishDate | 2023-06-01 |
| publisher | Future Science Ltd |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj.art-fbf481aee640489db6e1383d79b060a22023-07-25T10:35:25ZengFuture Science LtdBioTechniques0736-62051940-98182023-06-0174631733210.2144/btn-2022-0127PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNAYasunori Nishida0Kotetsu Kayama1Taichi Endoh2Kiwamu Hanazono3Gerry Amor Camer4Daiji Endoh51Department of Radiation Biology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 069-8501, Japan1Department of Radiation Biology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 069-8501, Japan1Department of Radiation Biology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 069-8501, Japan1Department of Radiation Biology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 069-8501, Japan1Department of Radiation Biology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 069-8501, Japan1Department of Radiation Biology, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, 069-8501, JapanWe formulated a method to synthesize 1 kbp DNA fragments using ‘oligomer unidirectional joining method’ via asymmetric extension supported by a simulator for oligonucleotide extension (AESOE). In this study, trials were conducted on 41 sets of different genomic pieces of ten flaviviral genomes, and 31 bacterial 16s rRNA fragments with sizes ranging from 500 bases to 1.0 kbp. Synthetic gene production was found to be successful in all those sets. The synthesis method has three steps: the first step is a seven-linked AESOE, the second step is the linking of the 400-base fragments from the first step, and the third step is the final amplification. Our present approach is highly reproducible and may no longer require optimization of oligomer design.https://www.future-science.com/doi/10.2144/btn-2022-0127AESOEoptimization-free oligomer joiningsynthetic genesunisense joining oligomers |
| spellingShingle | Yasunori Nishida Kotetsu Kayama Taichi Endoh Kiwamu Hanazono Gerry Amor Camer Daiji Endoh PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA BioTechniques AESOE optimization-free oligomer joining synthetic genes unisense joining oligomers |
| title | PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA |
| title_full | PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA |
| title_fullStr | PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA |
| title_full_unstemmed | PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA |
| title_short | PCR-based gene synthesis with overlapping unisense-oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsDNA |
| title_sort | pcr based gene synthesis with overlapping unisense oligomers asymmetric extension supported by a simulator for oligonucleotide extension achieved 1 kbp dsdna |
| topic | AESOE optimization-free oligomer joining synthetic genes unisense joining oligomers |
| url | https://www.future-science.com/doi/10.2144/btn-2022-0127 |
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