Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing
A number of questions arise when choosing methods for experiments related to next-generation sequencing. On the one hand, while working with RNA extraction, added reagents and their residues can often inhibit sensitive chemicals with which the sequential synthesis is carried out for the sequencing....
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Bashkir State Medical University
2019-04-01
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| Series: | Креативная хирургия и онкология |
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| Online Access: | https://www.surgonco.ru/jour/article/view/367 |
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| author | O. A. Beylerli A. T Beylerli I. F. Garaev |
| author_facet | O. A. Beylerli A. T Beylerli I. F. Garaev |
| author_sort | O. A. Beylerli |
| collection | DOAJ |
| description | A number of questions arise when choosing methods for experiments related to next-generation sequencing. On the one hand, while working with RNA extraction, added reagents and their residues can often inhibit sensitive chemicals with which the sequential synthesis is carried out for the sequencing. On the other hand, processing the same data using different software for the analysis can also impact on the sequencing results. This paper will present the step by step procedure for the preparation of samples taken from human biological fluids for subsequent sequencing of small RNAs, small noncoding RNAs in particular. Regarding the methods of extraction or isolation of RNAs, we found that low RNA yield can be improved significantly by following the isolation method for total RNA and its fractions included in Ambion’s MirVana PARIS kit, but only if using a special approach and modifying the organic extraction step. Compared to others, the methods supplied with commercially available kits at the time of researching this paper require only one organic extraction. This simple but, as it turned out, very useful modification makes it possible to access previously unavailable material. Potential advantages of this modification include a more complete profiling of small non-coding RNAs and a broader access to small sample volumes, as a rule, access to human biological fluids which can be prepared for RNA sequencing on the Illumina platform. |
| format | Article |
| id | doaj-art-c0ea6b3a69b140d081de67f41bbc8133 |
| institution | DOAJ |
| issn | 2076-3093 2307-0501 |
| language | English |
| publishDate | 2019-04-01 |
| publisher | Bashkir State Medical University |
| record_format | Article |
| series | Креативная хирургия и онкология |
| spelling | doaj-art-c0ea6b3a69b140d081de67f41bbc81332025-08-20T03:19:20ZengBashkir State Medical UniversityКреативная хирургия и онкология2076-30932307-05012019-04-0191808610.24060/2076-3093-2019-9-1-80-86316Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation SequencingO. A. Beylerli0A. T Beylerli1I. F. Garaev2Bashkir State Medical UniversityBashkir State Medical UniversityBashkir State Medical UniversityA number of questions arise when choosing methods for experiments related to next-generation sequencing. On the one hand, while working with RNA extraction, added reagents and their residues can often inhibit sensitive chemicals with which the sequential synthesis is carried out for the sequencing. On the other hand, processing the same data using different software for the analysis can also impact on the sequencing results. This paper will present the step by step procedure for the preparation of samples taken from human biological fluids for subsequent sequencing of small RNAs, small noncoding RNAs in particular. Regarding the methods of extraction or isolation of RNAs, we found that low RNA yield can be improved significantly by following the isolation method for total RNA and its fractions included in Ambion’s MirVana PARIS kit, but only if using a special approach and modifying the organic extraction step. Compared to others, the methods supplied with commercially available kits at the time of researching this paper require only one organic extraction. This simple but, as it turned out, very useful modification makes it possible to access previously unavailable material. Potential advantages of this modification include a more complete profiling of small non-coding RNAs and a broader access to small sample volumes, as a rule, access to human biological fluids which can be prepared for RNA sequencing on the Illumina platform.https://www.surgonco.ru/jour/article/view/367rna extractionsmall rnanext-generation sequencingcerebrospinal fluidbiological fluids |
| spellingShingle | O. A. Beylerli A. T Beylerli I. F. Garaev Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing Креативная хирургия и онкология rna extraction small rna next-generation sequencing cerebrospinal fluid biological fluids |
| title | Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing |
| title_full | Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing |
| title_fullStr | Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing |
| title_full_unstemmed | Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing |
| title_short | Extracting Small RNAs from Human Biological Fluids for Subsequent Next-Generation Sequencing |
| title_sort | extracting small rnas from human biological fluids for subsequent next generation sequencing |
| topic | rna extraction small rna next-generation sequencing cerebrospinal fluid biological fluids |
| url | https://www.surgonco.ru/jour/article/view/367 |
| work_keys_str_mv | AT oabeylerli extractingsmallrnasfromhumanbiologicalfluidsforsubsequentnextgenerationsequencing AT atbeylerli extractingsmallrnasfromhumanbiologicalfluidsforsubsequentnextgenerationsequencing AT ifgaraev extractingsmallrnasfromhumanbiologicalfluidsforsubsequentnextgenerationsequencing |