GSM5222644: ccRCC1, Right clear cell renal cell carcinoma Homogenate; Homo sapiens; RNA-Seq
We used 10x Genomics Chromium Single Cell 3′ Reagents Kit v3 user guide   to perform single cell suspension which were prepared before. Briefly, the  single-cell samples were passed through a 40μm cells strainer and counted using  a hemocytometer with trypan blue. Subsequently, the appropriate volume for each sample was calculated to  recover 10,000 cells. With the 10x protocol, we added the single cells, the Gel Beads and the  oils onto the 10x Genomics single-cell-A chip and run this machine. After  droplet generation, samples were transferred onto a PCR tube and reverse  transcription was performed using T100 Thermal Cycler (BIO-RAD). After the  reverse transcription, cDNA was recovered using Recovery Agent provided by 10x genomics  followed by a Silane DynaBead clean-up as outlined in the user guide. Purified  cDNA was amplified for 10 cycles before being cleaned up by using SPRIselect  beads. The samples were detected by Qubit2.0 Fluorometer (invitrogen) to  determine cDNA concentration. 10X Genomics Single Cell 3' v3 Reagent Kit
GEO
GSM5222644