Namocell Single Cell Separator Application---Single Cell Sequencing

Namocell & Chan Zuckerburg Biohub Joint Development of Single Cell Sequencing Applications
In November 2018, Namocell teamed up with CZ-Biohub (Chan Zuckerburg Biohub) to make new attempts in single-cell sequencing. CZ-Biohub uses the Namocell single cell separator to sort out the B cells of interest and sequence them for single cell, which is an important step for the discovery of new antibody drugs.
Single-cell RNA sequencing (scRNA-seq) is widely used as a whole-genome sequencing method to identify cell types and cell variants at the single-cell level. The Namocell single cell separator can sort individual live B cells by cell labeling CD27 and a cell active dye. By analyzing the expression of isolated single-cell genes and comparing resting memory B cells, gene expression differences can be studied from a single cell level.
Figure 1. Human immature memory B cells were cultured in batches for 6 days in mCD40L-3T3. Cells were harvested and stained with CD27-PE and CellTracker Green. PE/FITC double positive cells were screened by Namocell single cell separator, single cells were sorted into 96 well PCR plates, and 4 ul/well of lysate was pre-plated in the pre-PCR plate. The ratio of single cell sorting was 97% by cDNA synthesis. Next, for amplify V H and V L, an antibody variable region (A, n = 8) Preparation of specific genes or whole transcriptome libraries; Generation Sequencing by acquisition sequence, with reference to resting memory cells expression of antibody B BCR Profile The group was compared and analyzed (B); the difference of B cell activation genes (C); the overall gene rise and fall rule (D). Only one set per well BCR transcript, as well as V H and V L, determined by the Sanger sequencing quality, fully described only a cell sorting into PCR tubes.
The above experimental data determined the efficiency of the sorted single cells, and also confirmed that the B cells were determined to be activated by pre-experimental treatment and gating criteria. More importantly, cDNA from sorted single cells is suitable for subsequent single cell transcriptome library preparation and full length PCR of immunoglobulin light heavy chain variable regions. The Namocell Single Cell Separator provides a new desktop single cell sorting platform that seamlessly interfaces with downstream single cell sequencing assays.

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