Scholar
Marketplace
Lab
Feedback
$
English
Sign In
Home
Search
My boards
Insights
Collaborators
Reader
Blog
Help
Home
Search
My boards
Insights
Collaborators
Reader
Blog
Help
Isolation of Plant Root Nuclei for Single Cell RNA Sequencing
Translate
Afrikaans
Albanian
Amharic
Arabic
Armenian
Azerbaijani
Basque
Belarusian
Bengali
Bosnian
Български
Catalan
Cebuano
中文(简体)
中文(繁體)
Corsican
Czech
Danish
Dutch
English
Esperanto
Estonian
Finnish
Français
Georgian
Deutsch
Greek
Haitian Creole
Hindi
Hungarian
Icelandic
Indonesian
Irish
Italian
Japanese
Kazakh
Khmer
Korean
Kyrgyz
Lao
Latin
Latvian
Lithuanian
Luxembourgish
Macedonian
Malay
Maltese
Maori
Mongolian
Myanmar (Burmese)
Nepali
Norwegian
Pashto
Persian
Polish
Portuguese
Romanian
Russian
Samoan
Serbian
Sinhala
Slovak
Slovenian
Somali
Spanish
Swahili
Swedish
Tajik
Tamil
Thai
Turkish
Ukrainian
Urdu
Uzbek
Vietnamese
Xhosa
Zulu
Dirk Anderson
Sandra Thibivilliers
Marc Libault
Journal:
2020/October
-
Current protocols in plant biology
PUBMED:
33034428
DOI:
10.1002/cppb.20120
Abstract:
The characterization of the transcriptional similarities and differences existing between plant cells and cell types is important to better understand the biology of each cell composing the plant, to reveal new molecular mechanisms controlling gene activity, and to ultimately implement meaningful strategies to enhance plant cell biology. To gain a deeper understanding of the regulation of plant gene activity, the individual transcriptome of each plant cell needs to be established. Until recently, single cell approaches were mostly limited to bulk transcriptomic studies on selected cell types. Accessing specific cell types required the development of labor-intensive strategies. Recently, single cell sequencing strategies were successfully applied on isolated Arabidopsis thaliana root protoplasts. However, this strategy relies on the successful isolation of viable protoplasts upon the optimization of the enzymatic cocktails required to digest the cell wall and on the compatibility of fragile plant protoplasts with the use of microfluidic systems to generate single cell transcriptomic libraries. To overcome these difficulties, we present a simple and fast alternative strategy: the isolation and use of plant nuclei to access meaningful transcriptomic information from plant cells. This protocol was specifically developed to enable the use of the plant nuclei with 10× Genomics' Chromium technology partitions technology. Briefly, the plant nuclei are released from the root by chopping into a nuclei isolation buffer before purification by filtration then nuclei sorting. Upon sorting, the nuclei are resuspended in a low divalent ion buffer compatible with the Chromium technology in order to create single nuclei ribonucleic acid-sequencing libraries (sNucRNA-seq). © 2020 Wiley Periodicals LLC. Basic Protocol 1: Arabidopsis seed sterilization and planting Basic Protocol 2: Nuclei isolation from Arabidopsis roots Basic Protocol 3: Fluorescent-activated nuclei sorting (FANS) purification Support Protocol: Estimation of nuclei density using Countess II automated cell counter Alternate Protocol 1: Proper growth conditions for Medicago truncatula and Sorghum bicolor Alternate Protocol 2: Estimation of nuclei density using sNucRNA-seq technology.
Keywords:
FANS; nuclei; roots; sNucRNA-seq; scRNA-seq; single cell RNA-sequencing; single nuclei RNA-sequencing.
Open in
PUBMED
|
DOI
|
Google Scholar
|
Wikipedia
Relations:
Citations
(3)
Chemicals
(1)
Organisms
(3)
Processes
(2)
Anatomy
(4)
Similar articles
Articles by the same authors
Discussion board
Collaboration tool especially designed for Life Science professionals.
Drag-and-drop any entity to your messages.
Learn More
Search and select entities to see details here. Details will include lists of related entities (via publication). Explore related entities further.
Learn More