ScRNA-seq libraries were constructed according to the “Single Cell 3' Reagent Kits v2 User Guide” (10X Genomics). Generally, single-cell population was barcoded, and barcoded cDNA was prepared inside each cell by reverse transcription. Cell lysis followed, and then cDNA library was achieved through a released barcoded cDNA amplification. Following fragmentation, end repair, and addition of a single A base, double-sided size selection was used to isolate cDNA around 200 bp. Further adaptor ligation, sample index PCR amplification, and another double-sided size selection, the final 300∼600 bp DNA sequencing library was constructed and sequenced on Illumina HiSeq by Novogene (Chula Vista, CA). The RNA-seq analysis methodology was published previously [40 (link)]. Heat maps were generated by CLC Genomics Workbench (QIAGEN Inc.).
Single cell 3 reagent kits v2 user guide
Manufactured by 10x Genomics
The Single Cell 3' Reagent Kits v2 User Guide provides instructions for using the reagents required to prepare single-cell samples for gene expression analysis using 10x Genomics' sequencing platforms. The guide covers sample preparation, library construction, and sequencing steps.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using single cell 3 reagent kits v2 user guide
1
Single-cell RNA-seq library construction
2
Single-Cell Viability Profiling and Sequencing
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For each sample, an aliquot of cells was assessed for viability with calcein-AM and ethidium-homodimer1 (P/N L3224 ThermoFisher Scientific). Single-cell capture was performed following the Single Cell 3′ Reagent Kits v2 User Guide (CG0052 10X Genomics). The viability was >70% each sample. Cell barcodes and unique molecular identifiers (UMI) barcodes were demultiplexed and single-end reads aligned to the reference genome, GRCh38, using the CellRanger pipeline (10X Genomics).
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!