简介：
Overview
This protocol outlines a reliable and optimized method for isolating nuclei from solid tumor specimens for multi-om sequencing using the 10x Genomics platform. It includes recommendations for tissue dissociation conditions and cryopreservation of single-cell suspensions.
Key Study Components
Area of Science
- Neuroscience
- Oncology
- Genomics
Background
- Understanding tumor biology is crucial for improving patient outcomes.
- Analyzing heterogeneous cells in the tumor microenvironment is increasingly important.
- Multi-om sequencing allows for paired acquisition of single-cell RNA and ATAC sequencing.
- The quality of data is highly dependent on experimental conditions and biological material.
Purpose of Study
- To provide a reliable protocol for isolating nuclei from solid tumors.
- To enhance the quality of multi-om sequencing data.
- To facilitate the analysis of tumor microenvironments.
Methods Used
- Isolation of nuclei from solid tumor specimens.
- Use of fresh or frozen single-cell suspensions.
- Recommendations for tissue dissociation conditions.
- Cryopreservation techniques for single-cell suspensions.
Main Results
- Optimized approach for nuclei isolation was established.
- Reliable results obtained from both fresh and frozen samples.
- Improved data quality for multi-om sequencing.
- Guidelines for tissue processing and cryopreservation were provided.
Conclusions
- The protocol enhances the reliability of nuclei isolation from tumors.
- It supports the advancement of multi-om sequencing techniques.
- Future studies can benefit from improved experimental conditions.
What is multi-om sequencing?
Multi-om sequencing allows for the simultaneous analysis of different molecular layers, such as RNA and chromatin accessibility, from the same cell.
Why is nuclei isolation important?
Isolating nuclei is crucial for studying the genetic and epigenetic landscape of cells within tumors, which can inform treatment strategies.
Can this protocol be used with frozen samples?
Yes, the protocol is optimized for both fresh and frozen single-cell suspensions.
What are the key recommendations for tissue dissociation?
The protocol provides specific conditions for tissue dissociation to ensure high-quality nuclei isolation.
How does cryopreservation affect sample quality?
Proper cryopreservation techniques help maintain the integrity of single-cell suspensions for reliable downstream analyses.
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