title: GSM5640072: Control; Mus musculus; RNA-Seq
library_construction_protocol: Samples were washed twice in PBS (Life Technologies) plus 0.04% BSA (Sigma‐Aldrich). Each wash was performed with a 5-min centrifugation at 330 g and resuspension in 1 ml PBS plus 0.04% BSA. Sample viability was assessed using trypan blue (Thermo Fisher) and a hemocytometer (Thermo Fisher), and the appropriate volume for each sample was calculated. After droplet generation (the generation of GEMs with RT primers), samples were transferred onto a prechilled 96-well plate (Eppendorf), and reverse transcription was performed. Next, post GEM-RT was recovered using Recovery Agent provided by 10x Genomics followed by DynaBead cleanup cDNA libraries were prepared as outlined by the Single Cell 3′ Reagent kit v2 user guide with appropriate modifications to the PCR cycles based on the calculated cDNA concentration (as recommended by 10x Genomics).