Detailed In Situ Hybridization Protocol

ISH was performed on 16-μm sections as described previously.16 (link),17 (link) Sense and antisense riboprobes were generated against full-length CASK IMAGE Clone (catalog number MMM1013-202761641; Dharmacon, Lafayette, CO, USA). Riboprobes were synthesized using digoxigenin (DIG; Roche, Mannheim, Germany) and the MAXI-Script In Vitro Transcription Kit (Ambion, Austin, TX, USA). Probes were hydrolyzed to approximately 500 base pairs. Images were obtained on a Zeiss LSM700 confocal microscope (Carl Zeiss Meditec, Jena, Germany).

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Kerr A., Patel P.A., LaConte L.E., Liang C., Chen C.K., Shah V., Fox M.A, & Mukherjee K. (2019). Non-Cell Autonomous Roles for CASK in Optic Nerve Hypoplasia. Investigative Ophthalmology & Visual Science, 60(10), 3584-3594.

Publication 2019

MAXI-Script In Vitro Transcription Kit Zeiss LSM700 confocal microscope

Austin Clone Confocal microscope Digoxigenin Transcription

Corresponding Organization : Carilion Clinic

Other organizations : Baylor College of Medicine, Texas Children's Hospital

Top 5 similar protocols

Variable analysis

independent variables

Sense and antisense riboprobes

dependent variables

Expression of CASK
Images obtained on a Zeiss LSM700 confocal microscope

control variables

16-μm sections
Full-length CASK IMAGE Clone (catalog number MMM1013-202761641; Dharmacon, Lafayette, CO, USA)
Digoxigenin (DIG; Roche, Mannheim, Germany)
MAXI-Script In Vitro Transcription Kit (Ambion, Austin, TX, USA)

controls

Sense and antisense riboprobes (positive and negative controls)

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!