Jun 03, 2020
Digoxigenin Labeled In Vitro Transcription and Ribogreen Quantification
- Allen Institute for Brain Science1
- 1Allen Institute
- BICCN / BICAN
- Allen Institute for Brain Science
Protocol Citation: Allen Institute for Brain Science 2020. Digoxigenin Labeled In Vitro Transcription and Ribogreen Quantification. protocols.io https://dx.doi.org/10.17504/protocols.io.bg4vjyw6
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: June 03, 2020
Last Modified: March 16, 2021
Protocol Integer ID: 37749
Keywords: MB0002, IVT, in vitro transcription, DIG-labeled RNA,
Abstract
This protocol is used for in vitro transcription (IVT) reaction generating DIG-labeled anti-sense (AS) RNA from polymerase chain reaction (PCR) product for in situ hybridization (ISH), as well as fluorescent quantification of products using Ribogreen immediately following. This protocol is written for 96 reactions but can also be used for single reactions or in multiples of 96 by adjusting the amount of materials and equipment appropriately.
Note: Research reported in this publication was supported by the National Institute Of Mental Health of the National Institutes of Health under Award Number U19MH114830. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.
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