The liver was isolated from mice 4 days after the hydrodynamic injection of 20 μg pUC19 or pHBV1.3. A single-cell suspension of the mouse liver was prepared by the two-step enzyme digestion procedure as described above. Most of parenchymal cells were removed. Cell samples were prepared by following the 10× Genomics Single Cell 3′ v3 Reagent Kit user guide. Briefly, cell samples were washed twice with PBS plus 0.04% BSA, resuspended in the same wash solution and then filtered with a 40-μm cell strainer. The cell viability was assessed using the Trypan Blue exclusion assay (Thermo Fisher) and counted using a hemocytometer. Following cell counting, the appropriate volume for each cell sample was calculated for a target capture of 5000 cells with a cell concentration of 1000 cells/μl. Cells were loaded onto the 10x Genomics single-cell-G chip, and the Chromium Single Cell 3′Reagent Kit v3 was used for the preparation of the cDNA according to the manufacturer’s protocol. Following the library preparation and quantitation, the libraries were sequenced on the Illumina NextSeq 6000 platform.
Genomics single cell g chip
Manufactured by 10x Genomics
The 10x Genomics single-cell-G chip is a microfluidic device designed for single-cell analysis. The chip allows for the encapsulation of individual cells in gel beads, enabling the isolation and barcoding of genetic material from each cell. This technology facilitates the study of gene expression patterns and cellular heterogeneity at the single-cell level.
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2 protocols using genomics single cell g chip
1
Single-Cell RNA-Seq of Mouse Liver after Hydrodynamic Injection
2
Single-Cell RNA-Seq of Mouse Liver after Hydrodynamic Injection
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The liver was isolated from mice 4 days after the hydrodynamic injection of 20 μg pUC19 or pHBV1.3. A single-cell suspension of the mouse liver was prepared by the two-step enzyme digestion procedure as described above. Most of parenchymal cells were removed. Cell samples were prepared by following the 10× Genomics Single Cell 3′ v3 Reagent Kit user guide. Briefly, cell samples were washed twice with PBS plus 0.04% BSA, resuspended in the same wash solution and then filtered with a 40-μm cell strainer. The cell viability was assessed using the Trypan Blue exclusion assay (Thermo Fisher) and counted using a hemocytometer. Following cell counting, the appropriate volume for each cell sample was calculated for a target capture of 5000 cells with a cell concentration of 1000 cells/μl. Cells were loaded onto the 10x Genomics single-cell-G chip, and the Chromium Single Cell 3′Reagent Kit v3 was used for the preparation of the cDNA according to the manufacturer’s protocol. Following the library preparation and quantitation, the libraries were sequenced on the Illumina NextSeq 6000 platform.
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