RT-PCR amplification of mRNA from single brain neurospheres.
J Neurosci Methods
; 96(1): 57-61, 2000 Mar 01.
Article
em En
| MEDLINE
| ID: mdl-10704671
ABSTRACT
A method is described that allows cDNA production from individual brain cell clones or 'neurospheres'. These culture-generated spheres of stem, progenitor, and differentiated cells have been the focus of interest because they represent an in vitro model of neurogenesis. However, because neurospheres are somewhat resistant, in part due to their enclosure by a dense extracellular matrix, to methods attempting to disrupt them and isolate nucleic acids, there is a need for new technology that affords the simple and efficient RT-PCR for studies of neural gene expression and discovery. A method is described here that uses sonication and an all-in-one approach for the construction of cDNA from single neurospheres. The generation of cDNA from individual adult brain stem/progenitor cell neurospheres is useful for future studies of neurogenic gene expression.
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Bases de dados:
MEDLINE
Assunto principal:
Células-Tronco
/
Encéfalo
/
RNA Mensageiro
/
Técnicas de Cultura de Células
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Limite:
Animals
Idioma:
En
Revista:
J Neurosci Methods
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
Estados Unidos