Bone marrow derived mouse DCs were prepared as previously described18 (link) and stimulated with pathogenic stimuli for specified time periods. The C1 Single-Cell Auto Prep System (Fluidigm) was used to perform SMARTer (Clontech) whole transcriptome amplification (WTA)15 ,16 (link),19 on up to 96 individual cells. WTA products were then converted to Illumina sequencing libraries using Nextera XT (Illumina)15 . RNA-Seq libraries were also made from 10,000 cells from each parent population (population control). Each sample was sequenced on an Illumina HiSeq 2000 or 2500, and expression estimates (transcripts per million; TPM) for all UCSC-annotated mouse genes were calculated using RSEM36 (link). Data was further analyzed as described in the SI . Additional experiments were performed using RNA-FISH (Panomics), “on-chip” isolated stimulation, knockout mice, secretion blockers (GolgiPlug, BD Biosciences), protein synthesis blockers (Cycloheximide, Sigma), and recombinant cytokines. Full Methods and any associated references are provided in SI . Data are deposited in GEO under accession number GSE48968.
Rna fish
Manufactured by Panomics
RNA-FISH is a laboratory technique used to visualize and localize specific RNA molecules within cells. It allows for the detection and analysis of gene expression patterns by labeling and detecting target RNA sequences using fluorescent probes.
Automatically generated - may contain errors
2 protocols using rna fish
1
Single-Cell RNA-Seq of Mouse Dendritic Cells
2
Single-Cell RNA-Seq of Mouse Dendritic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bone marrow derived mouse DCs were prepared as previously described18 (link) and stimulated with pathogenic stimuli for specified time periods. The C1 Single-Cell Auto Prep System (Fluidigm) was used to perform SMARTer (Clontech) whole transcriptome amplification (WTA)15 ,16 (link),19 on up to 96 individual cells. WTA products were then converted to Illumina sequencing libraries using Nextera XT (Illumina)15 . RNA-Seq libraries were also made from 10,000 cells from each parent population (population control). Each sample was sequenced on an Illumina HiSeq 2000 or 2500, and expression estimates (transcripts per million; TPM) for all UCSC-annotated mouse genes were calculated using RSEM36 (link). Data was further analyzed as described in the SI . Additional experiments were performed using RNA-FISH (Panomics), “on-chip” isolated stimulation, knockout mice, secretion blockers (GolgiPlug, BD Biosciences), protein synthesis blockers (Cycloheximide, Sigma), and recombinant cytokines. Full Methods and any associated references are provided in SI . Data are deposited in GEO under accession number GSE48968.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!