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Name Authors DOI Group Summary Associated Publications RRIDs used Affiliations External URL Version Publication Date Proper Citation Record Last Update
Phosphate Buffered Saline (PBS)
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bezgjf3w BICCN, Allen Institute for Brain Science This protocol is used to prepare Phosphate Buffered Saline (PBS). PBS (-) and PBS (+) which includes Tween are used as biologically neutral wash solutions in many processes including In Situ Hybridization (ISH), Immunohistochemistry (IHC) and Fixation.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. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Phosphate Buffered Saline (PBS). protocols.io https://dx.doi.org/10.17504/protocols.io.bezgjf3w 2021-04-15 09:16:09
PCR Prep from cDNA for IVT Reactions
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bcsfiwbn BICCN, Allen Institute for Brain Science This protocol describes PCR1 and PCR2 reactions beginning with cDNA to generate transcript templates ready for IVT reaction. This protocol is written for 192 (2x96) reactions but can also be used for fewer or more reactions by adjusting the amount of materials and equipment appropriately. Allen Institute 1 2020 Allen Institute for Brain Science 2020. PCR Prep from cDNA for IVT Reactions. protocols.io https://dx.doi.org/10.17504/protocols.io.bcsfiwbn 2021-04-15 09:16:09
Mouse Whole Cell Tissue Processing for 10x Genomics Platform
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.be2fjgbn BICCN, Allen Institute for Brain Science This protocol describes how to isolate tissue from various ROI’s of the mouse brain followed by preparation of a highly concentrated single cell suspension for transcriptomic profiling on 10x Genomics platform.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. Allen Institute 7 2020 Allen Institute for Brain Science 2020. Mouse Whole Cell Tissue Processing for 10x Genomics Platform. protocols.io https://dx.doi.org/10.17504/protocols.io.be2fjgbn 2021-04-15 09:16:13
Nextera XT at 0.2X On the Mantis
 
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ZengU19 BRAIN Grant DOI:10.17504/protocols.io.63rhgm6 BICCN, Allen Institute for Brain Science Protocol to generate Nextera libraries using 0.2x reagents in a 96-well PCR plate using the Formulatrix Mantis instrument. Allen Institute 1 2019 ZengU19 BRAIN Grant 2019. Nextera XT at 0.2X On the Mantis. protocols.io https://dx.doi.org/10.17504/protocols.io.63rhgm6 2021-04-15 09:16:13
Production of Crude AAV Virus Extract
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bdvwi67e BICCN, Allen Institute for Brain Science This protocol is used to produce crude preps of AAV of any serotype.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. Allen Institute 3 2020 Allen Institute for Brain Science 2020. Production of Crude AAV Virus Extract. protocols.io https://dx.doi.org/10.17504/protocols.io.bdvwi67e 2021-04-15 09:16:22
TissueCyte Troubleshooting Guide
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.beryjd7w BICCN, Allen Institute for Brain Science This protocol outlines strategies to solve common problems that occur when setting up the TissueCyte 1000 system for normal operation.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. Allen Institute 3 2020 Allen Institute for Brain Science 2020. TissueCyte Troubleshooting Guide. protocols.io https://dx.doi.org/10.17504/protocols.io.beryjd7w 2021-04-15 09:16:27
Slice Preparation with Tissue Dissociation - Mouse Protocol
 
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ZengU19 BRAIN grant DOI:10.17504/protocols.io.7dwhi7e BICCN, Allen Institute for Brain Science This protocol describes the procedure for transcardial perfusion of an adult mouse, followed by brain dissection, embedding, slicing, blockface imaging, ROI microdissection, and tissue dissociation. The output of this protocol is a cell suspension that can be sorted via FACS for single-cell transcriptional profiling. Allen Institute 1 2019 ZengU19 BRAIN grant 2019. Slice Preparation with Tissue Dissociation - Mouse Protocol. protocols.io https://dx.doi.org/10.17504/protocols.io.7dwhi7e 2021-04-15 09:16:25
Mouse Cardiac Perfusion Fixation and Brain Collection
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bd8vi9w6 BICCN, Allen Institute for Brain Science This protocol describes the procedures for intracardiac perfusion fixation of postnatal mice, including anesthesia, exsanguination, fixation, brain removal and post-fixation storage. 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. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Mouse Cardiac Perfusion Fixation and Brain Collection. protocols.io https://dx.doi.org/10.17504/protocols.io.bd8vi9w6 2021-04-15 09:16:19
Patch-Seq Recording and Extraction
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bcthiwj6 BICCN, Allen Institute for Brain Science This protocol describes the process to obtain electrophysiological recordings and cellular contents from neurons in postnatal mouse and/or human brain slices. Allen Institute 1 2020 Allen Institute for Brain Science 2020. Patch-Seq Recording and Extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.bcthiwj6 2021-04-15 09:16:32
TissueCyte Specimen Embedding Acrylamide Coembedding
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bdpvi5n6 BICCN, Allen Institute for Brain Science This protocol describes the preparation of a fixed mouse brain specimen bonded to an agarose block for blockface imaging and sectioning using the TissueCyte 1000 system.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. Allen Institute 2 2020 Allen Institute for Brain Science 2020. TissueCyte Specimen Embedding Acrylamide Coembedding. protocols.io https://dx.doi.org/10.17504/protocols.io.bdpvi5n6 2021-04-15 09:16:22
DAB Detection of Biocytin Labeled Tissue
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.be2cjgaw BICCN, Allen Institute for Brain Science This protocol describes the process for diaminobenzidine (DAB) detection of biocytin filled cells. This protocol is optimized for use with brain slices cut at 350 µm thick, in which cells are first filled with biocytin (i.e., post-electrophysiological recording), fixed in 4% PFA/2.5% glutaraldehyde, and transferred to PBS until ready to stain.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. Allen Institute 4 2020 Allen Institute for Brain Science 2020. DAB Detection of Biocytin Labeled Tissue. protocols.io https://dx.doi.org/10.17504/protocols.io.be2cjgaw 2021-04-15 09:16:39
DAB Detection of Biocytin Labeled Tissue
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bctbiwin BICCN, Allen Institute for Brain Science This protocol describes the process for diaminobenzidine (DAB) detection of biocytin filled cells. This protocol is optimized for use with brain slices cut at 350 µm thick, in which cells are first filled with biocytin (i.e., post-electrophysiological recording), fixed in 4% PFA/2.5% glutaraldehyde, and transferred to PBS until ready to stain. Allen Institute 1 2020 Allen Institute for Brain Science 2020. DAB Detection of Biocytin Labeled Tissue. protocols.io https://dx.doi.org/10.17504/protocols.io.bctbiwin 2021-04-15 09:16:45
Human Tissue Slicing and Dissections for Nuclear Isolations
 
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LeinU01 BRAIN Grant DOI:10.17504/protocols.io.7aehibe BICCN, Allen Institute for Brain Science This protocol describes the slicing and microdissection procedure on frozen post-mortem human brain tissue to be used to isolate nuclei for subsequent transcriptional profiling. Allen Institute 2 2019 LeinU01 BRAIN Grant 2019. Human Tissue Slicing and Dissections for Nuclear Isolations. protocols.io https://dx.doi.org/10.17504/protocols.io.7aehibe 2021-04-15 09:16:23
Nextera XT at 0.2X On the Mantis
 
Resource Report
Resource Website
Allen Institute for Brain Science DOI:10.17504/protocols.io.brdjm24n BICCN, Allen Institute for Brain Science Protocol to generate Nextera libraries using 0.2x reagents in a 96-well PCR plate using the Formulatrix Mantis instrument.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. Allen Institute 2 2021 Allen Institute for Brain Science 2021. Nextera XT at 0.2X On the Mantis. protocols.io https://dx.doi.org/10.17504/protocols.io.brdjm24n 2021-04-15 09:16:55
Mouse Whole Cell Tissue Processing for 10x Genomics Platform
 
Resource Report
Resource Website
Allen Institute for Brain Science DOI:10.17504/protocols.io.bc6bizan BICCN, Allen Institute for Brain Science This protocol describes how to isolate tissue from various ROI’s of the mouse brain followed by preparation of a highly concentrated single cell suspension for transcriptomic profiling on 10x Genomics platform. Allen Institute 3 2020 Allen Institute for Brain Science 2020. Mouse Whole Cell Tissue Processing for 10x Genomics Platform. protocols.io https://dx.doi.org/10.17504/protocols.io.bc6bizan 2021-04-15 09:16:30
Frozen Tissue Nuclei Extraction
 
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Carly Martin, Abdul Abdul, Charles Vanderburg, Naeem Nadaf, Ashley Feirrera, Evan Macosko DOI:10.17504/protocols.io.bbseinbe BICCN Protocol for extraction of nuclei from frozen tissue in preparation for single-nuclei sequencing (droplet-based/10X). This protocol is based strongly on a similar extraction protocol from the McCarroll lab. Broad Institute, Broad Institute, Broad Institute, Broad Institute, Broad Institute, Broad Institute 2 2020 Carly Martin, Abdul Abdul, Charles Vanderburg, Naeem Nadaf, Ashley Feirrera, Evan Macosko 2020. Frozen Tissue Nuclei Extraction. protocols.io https://dx.doi.org/10.17504/protocols.io.bbseinbe 2021-04-15 09:16:56
Tris Buffered Saline (TNT)
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.brdkm24w BICCN, Allen Institute for Brain Science This protocol is used to prepare Tris Buffered Saline (TNT). 10X TNT is the stock solution used to prepare 1X TNT, which equilibrates tissue and maintains pH in physiological range.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. Allen Institute 3 2021 Allen Institute for Brain Science 2021. Tris Buffered Saline (TNT). protocols.io https://dx.doi.org/10.17504/protocols.io.brdkm24w 2021-04-15 09:16:54
Coating Slides with Gelatin
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bf64jrgw BICCN, Allen Institute for Brain Science The protocol describes how 1x3 glass microscope slides are coated with gelatin to improve adherence of fixed adult mouse brain sections or other tissue to slides.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. Allen Institute 2 2020 Allen Institute for Brain Science 2020. Coating Slides with Gelatin. protocols.io https://dx.doi.org/10.17504/protocols.io.bf64jrgw 2021-04-15 09:16:57
50 mM Phosphate Buffer (with 0.02% sodium azide option)
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bg52jy8e BICCN, Allen Institute for Brain Science 50 mM Phosphate Buffer can be used as a general buffer solution for multiple applications. It is suitable for use as a buffer for specimen embedding, as part of brain preparation associated with TissueCyte 2P Serial Imaging, described in PF0315 TissueCyte Specimen Embedding Acrylamide Coembedding.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. Allen Institute 2 2020 Allen Institute for Brain Science 2020. 50 mM Phosphate Buffer (with 0.02% sodium azide option). protocols.io https://dx.doi.org/10.17504/protocols.io.bg52jy8e 2021-04-15 09:16:57
Pulling Pipettes using the DMZ-Zeitz Universal
 
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Allen Institute for Brain Science DOI:10.17504/protocols.io.bepzjdp6 BICCN, Allen Institute for Brain Science Protocol is used for the manufacture of glass pipettes for patch clamp electrophysiology using the DMZ-Zeitz Universal puller.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. Allen Institute 3 2020 Allen Institute for Brain Science 2020. Pulling Pipettes using the DMZ-Zeitz Universal. protocols.io https://dx.doi.org/10.17504/protocols.io.bepzjdp6 2021-04-15 09:16:40

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