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Protocols
Biochemistry
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Microbiology
Molecular Biology
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Articles and Issues
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Overview
Authored
(1)
Reviewed
(9)
Q&A
(2)
Feedback
(1)
LZ
Lei Zhang
Washington State University
Research fields
Plant science
In situ
Hybridization (ISH) in Preparasitic and Parasitic Stages of the Plant-parasitic Nematode
Meloidogyne
spp.
Authors:
Maëlle Jaouannet
,
Chinh-Nghia Nguyen
,
Michaël Quentin
,
Stéphanie Jaubert-Possamai
,
Marie-Noëlle Rosso
and
Bruno Favery
,
date:
03/20/2018,
view:
8435,
Q&A:
1
The spatio-temporal expression pattern of a gene provides important indications to better understand its biological function.
In situ
hybridization (ISH) uses a labeled complementary single-stranded RNA or DNA probe to localize gene transcripts in a whole organism, a whole organ or a section of tissue. We adapted the ISH technique to the plant parasite
Meloidogyne
spp. (root-knot nematode) to visualize RNAs both in free-living preparasitic juveniles and in parasitic stages settled in the plant tissues. We describe each step of the probe synthesis, digoxigenin (DIG) labeling, nematode extraction from plant tissue, and ISH procedure.
More >
Formaldehyde for fixing nematodes
[Feedback 1] Since the formaldehyde recommended in the protocol has been discontinued by Sigma, I tried a similar product from sigma: 37% formaldehyde solution (Sigma: 252549), and it worked well for the in situ hybridization.
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