scRNA-seq from 11dpp testis of mouse Mus musculus
50,000 nuclei were mixed with a modified nuclei buffer and reagents, incubated at 37C for 30 minutes, and then loaded into a BD Rhapsody Cartridge after adding sample buffer. Nuclei were sedimented, washed, and incubated with splint beads before being agitated and washed again. After lysis with DTT and proteinase K, tagmented DNA and mRNA were immobilized onto beads, followed by DNA ligation and reverse transcription using a mix containing dNTPs, RT buffer, and reverse transcriptase. For WTA library preparation, random priming and extension were performed, followed by purification with AMPure XP Beads. The WTA libraries were then amplified, indexed, and analyzed on a Bioanalyzer to verify DNA size and concentration.