Single-Cell RNA Sequencing Preamplification Protocol

Primers pool was prepared as described previously (18 (link)). Primers used are listed in Supplementary Table S4. Individual cells were picked up into 8-strip PCR tubes with 5 µL RT-PreAmp Master Mix (1.9 µL nuclease free water, 2.5 µL Reaction Mix, and 0.1 µL RT/Taq enzyme were mixed with 0.5 µL primers pool; Single Cell Sequence Specific Amplification Kit, Vazyme, China) by special Pasteur pipettes (Brand, Germany). Eight-strip PCR tubes were immediately frozen in -80°C refrigerator for 2 min. After brief centrifugation (300 g, 4°C, 3 min), tubes were immediately moved to PCR machine following kit instructions. After preamplification, samples were diluted 100-fold with double distilled water prior to qPCR.