Chapter title |
MicroRNA In Situ Hybridization on Whole-Mount Preimplantation Embryos
|
---|---|
Chapter number | 2 |
Book title |
In Situ Hybridization Protocols
|
Published in |
Methods in molecular biology, August 2014
|
DOI | 10.1007/978-1-4939-1459-3_2 |
Pubmed ID | |
Book ISBNs |
978-1-4939-1458-6, 978-1-4939-1459-3
|
Authors |
Goossens, Karen, Peelman, Luc, Van Soom, Ann, Karen Goossens, Luc Peelman, Ann Van Soom, Soom, Ann, Soom, Ann Van |
Abstract |
Whole-mount in situ hybridization (WISH) using antisense probes is widely used to visualize RNA sequences in embryos and to determine the precise site of expression in the different cells or tissues. The target sequence is hybridized with an antisense RNA probe, followed by visual or fluorescence detection to measure the site and level of expression. However, the detection of short RNA molecules is hampered by the reduced stringency of the probes for short transcripts. Here, we describe a procedure for WISH detection of short RNA molecules, like miRNAs, in mammalian preimplantation embryos using LNA-modified probes with high sensitivity and specificity. |
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