GSM8619749: C57_PBS_Day20_rep2; Mus musculus; RNA-Seq
RNA was harvested by using the lysis buffer provided by 10K Genomics, Shanghai, China. Low-input SMART-seq2 library construction was conducted by 10K Genomics, Shanghai, China. Initially, the lysate samples underwent vortexing and centrifugation, followed by heat treatment on a preheated PCR machine. Smart RT buffer and RT enzyme were then added, and the reverse transcription reaction was carried out according to the program. Subsequently, Smart Amp buffer-1 and Smart Amp enzyme-1 were added to the cDNA product obtained in the previous step, and the amplification reaction was conducted according to the procedure. A small amount of product was taken out for quality control via electrophoresis gel. Quantitative purification of cDNA was performed using a DNA purification kit. The purified cDNA samples were then mixed with TN5 buffer and TN5 enzyme-1, and the reaction was interrupted according to the program. After the reaction, Smart Amp buffer-2, Smart Amp enzyme-2, and index for library construction were added, followed by purification and sending for testing.