A nonradioactive whole-mount in situ hybridization protocol for Porphyra (Rhodophyta) gametophytic germlings |
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| Authors: | Yasuko Shimizu Yukihiro Kitade Naotsune Saga |
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| Affiliation: | (1) Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido, 041-8611, Japan |
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| Abstract: | The objective of this study was to develop a method for whole-mount in situ hybridization (WISH) by using elongation factor-1 (EF-1) riboprobes in the red alga Porphyra yezoensis Ueda. Several modifications to the general WISH protocol, such as use of a short-length probe, performing partial digestion of the cell wall, optimization of proteinase K concentration and additional washing steps after hybridization were essential to reduce non-specific staining and to obtain sufficient quality of data. This protocol made it possible to detect a specific signal as a positive control in WISH assays of P. yezoensis. |
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| Keywords: | EF-1 " target="_blank">gif" alt="agr" align="BASELINE" BORDER="0"> in situ hybridization mRNA localization Porphyra yezoensis Rhodophyta |
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