State of the art techniques have been developed to isolate and analyze cells from various tissues, aiming to capture their in vivo state. However, the majority of cell isolation protocols involve lengthy mechanical and enzymatic dissociation steps followed by flow cytometry, exposing cells to stress and disrupting their physiological niche. Focusing on adult skeletal muscle stem cells, we have developed a protocol that circumvents the impact of isolation procedures and captures cells in their native quiescent state. We show that current isolation protocols induce major transcriptional changes accompanied by specific histone modifications while having negligible effects on DNA methylation. In addition to proposing a protocol to avoid isolation-induced artifacts, our study reveals previously undetected quiescence and early activation genes of potential biological interest.
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产品
剪切技术
Automation
Chromatin studies
DNA methylation
- Bisulfite conversion
- Methylated DNA Immunoprecipitation
- Methylbinding domain protein
- Hydroxymethylated DNA Immunoprecipitation
Genome editing (CRISPR/Cas9)
Antibodies
- All antibodies
- Sample size antibodies
- ChIP-seq grade antibodies
- ChIP-grade antibodies
- Western Blot Antibodies
- DNA modifications
- RNA modifications
- CRISPR/Cas9 antibodies
- CUT&Tag Antibodies
NGS Library preparation
- Library preparation for ChIP-seq
- Library preparation for RNA sequencing
- Library preparation for DNA sequencing
Reagents
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- 公司
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