PacBio Iso-Seq library preparation and sequencing

Richard John Tiika; Jia Wei; Guangxin Cui; Yanjun Ma; Hongshan Yang; Huirong Duan

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PacBio Iso-Seq library preparation and sequencing

RT Richard John Tiika

JW Jia Wei

GC Guangxin Cui

YM Yanjun Ma

HY Hongshan Yang

HD Huirong Duan

This method is extracted from research article: BMC Plant Biol, Oct 2021

Transcriptome-wide characterization and functional analysis of Xyloglucan endo-transglycosylase/hydrolase (XTH) gene family of Salicornia europaea L. under salinity and drought stress

DOI: 10.1186/s12870-021-03269-y

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The SMRTbell™ Template Prep Kit 1.0-SPv3 (Pacific Biosciences, Menlo Park, CA, USA) was used to run the sequencing library on 1 μg total RNA from the mixed shoot and root sample. The Qubit 2.0 fluorometer (Life Technologies, Carlsbad, CA, USA) was used to check the final library amount and concentration. An Agilent 2100 Bioanalyzer was used to assess library size and purity (Agilent Technologies, Santa Clara, CA, USA). Magbead-loaded SMRTbell template was run on a PacBio Sequel instrument at Shanghai Oe Biotech Co., Ltd. according to Sequel Binding Kit 2.0 instructions (Pacific Bioscience, USA) for primer annealing and polymerase binding (Shanghai, China).

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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