NEXT GENERATION SEQUENCING METHOD OF ZOOBENTHOS CYTOCHROME OXIDASE SUBUNIT I GENE AND USE THEREOF
The present invention relates to the technical field of zoobenthos identification, and in particular to a next generation sequencing (NGS) method of a zoobenthos cytochrome oxidase subunit T (COT) gene and use thereof. The method 5 includes the following steps: Sl, amplifying the COT gene by polymer...
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Main Authors | , , , , , , , , , |
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Format | Patent |
Language | English |
Published |
19.07.2022
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Subjects | |
Online Access | Get full text |
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Summary: | The present invention relates to the technical field of zoobenthos identification, and in particular to a next generation sequencing (NGS) method of a zoobenthos cytochrome oxidase subunit T (COT) gene and use thereof. The method 5 includes the following steps: Sl, amplifying the COT gene by polymerase chain reaction (PCR) with a DNA sample of a mixed zoobenthos sample as a template; SZ, amplifying a PCR.product obtained in step Sl as a template by PCR, to obtain a PCR product as a DNA library, where an amplified primer pair includes a 10 pyroseguencing-NGS adapter and each sample corresponds to one pyroseguencing-NGS adapter; and S3, conducting guality detectioncm1aresultinglflwxlibraryy andselectingeagualified DNA_library to conduct emPCR.amplification.and.NGS. The method obtains a high-throughput full-length (~658 bp) COT gene 15 sequence with read length covering large benthic communities by all-at-once sequencing, providing a large taxonomic resolutionandavoidingfalsepositivesdueixnexcessivelyhigh throughput. |
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Bibliography: | Application Number: NL20202026403 |