GSM4455994: GS612_NT; Homo sapiens; RNA-Seq
We took our prepared solutions and created such concentration that was calculated to recover 10,000 cells. We used the 10x Genomics Chromium Single Cel 3' Reagents Kit v3. With the 10x protocol, we added the single cells, the Gel Beads and the oils onto the 10x Genomics single-cell-A chip and ran them on the machine. After the droplet generation, samples were transferred into a PCR tube and reverse transcription was performed. After the reverse transcription, cDNA was recovered using the Recovery Agent provided by 10x Genomics followed by a Silane DynaBead clean-up as outlined as described by the manufacturer. Purified cDNA was amplified for 12 cycles before being cleaned up by using SPRIselect beads. The samples were measured on an Agilent bioAnalyzer with a Highsense DNA chip machine to determine the cDNA concentration. The final qpcr library was quantified with the NEBnext library quant kit for Illumina on a Thermo Fast7500. RNA libraries were prepared for sequencing using standard Illumina protocols.