Single-nucleus RNA sequencing of cortical tissue from transgenic mice with neuron-specific SEC61A1 overexpression
Nucleus preparationThe nucleus was isolated using a Nucleus Isolation Kit (SHBIO 52009-10) as instructions. RNase inhibitors (Sigma 3335399001) were added to the reagents before use. The samples were cut and transferred to a 5 mL tube containing lysate, mixed and lysed for 3 min on ice, then filtered through a 40 m cell filter (Sigma BAH136800040). Nucleus count and viability was estimated using SeekMate Tinitan Fluorescence Cell Counter (SeekGene M002C) with AO/PI reagent. After staining with 0.4% trypan blue (Sangon Biotech E607320-0001), the nucleus was observed under a 40 microscope (Jiangnan Novel Optics XD-202). Subsequent experiments were performed if the nuclear envelope was intact and there were fewer impurities.