GSM8921752: SPF; Mus musculus; OTHER
Library preparation was conducted using the Nextera XT Library Prep Kit. Purified cDNA (1 ng) was diluted in RNase-free water to a total volume of 5 µL. Then, 10 µL of Tagment DNA buffer and 5 µL of Amplicon Tagment mix were added, and the mixture was incubated at 55°C for 5 minutes. Afterward, 5 µL of NT buffer was added and incubated at room temperature for 5 minutes. PCR master solution (15 µL of PCR master mix, 1 µL of 10 µM P5 primer, 1 µL of 10 µM indexed P7 primer, and 8 µL of RNase-free water) was added, and the PCR reaction was performed with the following program: 95°C for 30 seconds, followed by 12 cycles of 95°C for 10 seconds, 55°C for 30 seconds, 72°C for 30 seconds, and a final extension at 72°C for 5 minutes. The PCR product was purified using 0.6× Ampure XP beads to obtain the library.