Image from Google Jackets

A useful protocol for in situ RT-PCR on plant tissues

Tipo de material: TextoTextoSeries ; Cellular & Molecular Biology, 7, p.7-18, 2002Trabajos contenidos:
  • Urbanczyk-Wochniak, E
  • Filipecki, M
  • Przybecki, Z
Tema(s): Recursos en línea: Resumen: This study describes an effective method of in situ RT-PCR (RT-ISPCR)that was developed to localize gene expression in plant tissues. This RT-PCR technique was performed on sectioned tissues of female buds of the cucumber GY3 inbred line. The CUS1 gene, encoding the MADS-box type (agamus-like)protein, the expression pattern of which was described earlier [10], was used as a marker gene for optimisation of steps in the in situ RT-PCR inside the cells.For the identificat ion of RT-PCR products inside the cells of the female buds, they were fixed in FAA solution, embedded in Paraplast Plus and cut into7mm thick sections which were dewaxed by immersion in HistoClear and dehydrated with ethanol. They were washed in water, th en in 0.02M HCl, 2xSSC and PBS buffer. In the next step of tissue pretreatment, the sections were digested with
Tags from this library: No tags from this library for this title. Log in to add tags.
Star ratings
    Average rating: 0.0 (0 votes)
Holdings
Item type Current library Collection Call number Status Date due Barcode
Documentos solicitados Documentos solicitados CICY Documento préstamo interbibliotecario Ref1 B-15338 (Browse shelf(Opens below)) Available

This study describes an effective method of in situ RT-PCR (RT-ISPCR)that was developed to localize gene expression in plant tissues. This RT-PCR technique was performed on sectioned tissues of female buds of the cucumber GY3 inbred line. The CUS1 gene, encoding the MADS-box type (agamus-like)protein, the expression pattern of which was described earlier [10], was used as a marker gene for optimisation of steps in the in situ RT-PCR inside the cells.For the identificat ion of RT-PCR products inside the cells of the female buds, they were fixed in FAA solution, embedded in Paraplast Plus and cut into7mm thick sections which were dewaxed by immersion in HistoClear and dehydrated with ethanol. They were washed in water, th en in 0.02M HCl, 2xSSC and PBS buffer. In the next step of tissue pretreatment, the sections were digested with

There are no comments on this title.

to post a comment.