Isolation of Nuclei Tagged in Specific Cell Types (INTACT) in Arabidopsis

Sammanfattning

Many functionally distinct plant tissues have relatively low numbers of cells that are embedded within complex tissues. For example, the shoot apical meristem (SAM) consists of a small population of pluripotent stem cells surrounded by developing leaves and/or flowers at the growing tip of the plant. It is technically challenging to collect enough high-quality SAM samples for molecular analyses. Isolation of Nuclei Tagged in specific Cell Types (INTACT) is an easily reproducible method that allows the enrichment of biotin-tagged cell-type-specific nuclei from the total nuclei pool using biotin-streptavidin affinity purification. Here, we provide a detailed INTACT protocol for isolating nuclei from the Arabidopsis SAM. One can also adapt this protocol to isolate nuclei from other tissues and cell types for investigating tissue/cell-type-specific transcriptome and epigenome and their changes during developmental programs at a high spatiotemporal resolution. Furthermore, due to its low cost and simple procedures, INTACT can be conducted in any standard molecular laboratory.

Nyckelord

Arabidopsis; Biotinylation; Cell-type-specific promoter; in situ streptavidin histochemistry; INTACT; low-input RT-qPCR; Shoot apical meristem

Publicerad i

Methods in Molecular Biology
2023, volym: 2686, nummer: 2686, sidor: 313-328
Titel: Flower Development : Methods and Protocols
Utgivare: Humana Press Inc.

SLU författare

• Schmid, Markus

  • Institutionen för växtbiologi, Sveriges lantbruksuniversitet
    
  • Umeå Universitet

UKÄ forskningsämne

Cellbiologi
Botanik

Publikationens identifierare

• DOI: https://doi.org/10.1007/978-1-0716-3299-4_16
• ISBN: 978-1-0716-3298-7
• eISBN: 978-1-0716-3299-4

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/129516