A systematic comparison of protocols for recovery of high-quality rna from human islets extracted by laser capture microdissection

Teresa Mezza, Andrea Giaccari, Chiara Maria Assunta Cefalo, Rohit N. Kulkarni

Risultato della ricerca: Contributo in rivistaArticolo in rivista

Abstract

The isolation of high-quality RNA from endocrine pancreas sections represents a consider-able challenge largely due to the high ribonuclease levels. Laser capture microdissection (LCM) of mammalian islets, in association with RNA extraction protocols, has emerged as a feasible approach to characterizing their genetic and proteomic profiles. However, a validated protocol to obtain high-quality RNA from LCM-derived human pancreas specimens that is appropriate for next-generation sequencing analysis is still lacking. In this study, we applied four methods (Picopure extraction kit, Qiazol protocol, Qiazol + Clean-up kit, and RNeasy Microkit + Carrier) to extract RNA from human islets obtained from both non-diabetic individuals and patients with type 2 diabetes who had undergone partial pancreatectomy, as well as handpicked islets from both non-diabetic and diabetic organ donors. The yield and purity of total RNA were determined by 260/280 absorbance using Nanodrop 100 and the RNA integrity number with a bioanalyzer. The results indicated that among the four methods, the RNeasy MicroKit + Carrier (Qiagen) provides the highest yield and purity.
Lingua originaleEnglish
pagine (da-a)625-632
Numero di pagine8
RivistaBiomolecules
Volume11
DOI
Stato di pubblicazionePubblicato - 2021

Keywords

  • Human islets
  • Laser capture microdissection
  • RNA extraction

Fingerprint Entra nei temi di ricerca di 'A systematic comparison of protocols for recovery of high-quality rna from human islets extracted by laser capture microdissection'. Insieme formano una fingerprint unica.

Cita questo