title: GSM5471468: healthy mouse left ventricle1; Mus musculus; RNA-Seq
library_construction_protocol: After collecting left ventricular region of normal healthy hearts, the tissue was digested with enzymes (2 mg/mL Collagenase IV [Worthington Biochemical], 1.2 units/mL Dispase II [Sigma-Aldrich]) for 15 minutes and used for subsequent single-cell sequencing. Then cDNA fragments were 10× labeled. Next, gel beads containing barcode information were first combined with a mixture of cells and enzymes, and then surrounded by oil surfactant droplets located in the microfluidic "double cross" connection. The droplet was very small and can only hold one gel bead. After the droplet flowed into the reservoir and collected, the gel bead dissolved and released the primer sequence, the cDNA fragment was reverse transcribed, and the cDNA was used as a template for PCR amplification. The products containing barcode information in each droplet were mixed to construct a sequencing library. Then, the Biorupter ultrasonic disrupter was used to break the cDNA into fragments of about 200-300bp, the sequencing adapter P5 was added and sequencing primer R1 and other traditional second-generation sequencing library construction process, and finally perform PCR amplification to obtain the DNA library.