GSM9201732: black ball 1; Mus musculus; RNA-Seq
Single-cell amplification was performed using SMART-Seq® HT Kit (Takara, USA) according to the manufacturer's instructions. A total of 18 “black ball-like” cells from 9 mice (3 mice/group) were lysed in a buffer containing free dNTP and oligo(dT) oligonucleotides. The RNA fragments were copied into the first strand cDNA using reverse transcriptase and several template-free C nucleotides. The oligonucleotide primers were hybridized with poly(C) to form the second strand cDNA. cDNA was purified and enriched using PCR. The cDNA libraries were constructed using the KAPA Hyper Prep Kit (KAPA, USA) according to the manufacturer's instructions. Purified libraries were quantified and validated using a Qubit® 3.0 Fluorometer and an Agilent 2100 bioanalyzer to confirm the insert size and calculate the mole concentration. The RNA fragments were copied into the first strand cDNA using reverse transcriptase and several template-free C nucleotides. The oligonucleotide primers were hybridized with poly(C) to form the second strand cDNA. cDNA was purified and enriched using PCR. The cluster was generated by cBot after the library was diluted to 10 pM and sequenced on the Illumina NovaSeq 6000 platform (Illumina, USA).