RNA-Seq of Mus musculus: 7-day-old male or female suckling mice small intestine
The number of biological replicates for each group of samples is three. The main experimental steps are as follows: First, extract total RNA from samples. Then, detect the RNA and if it doesn't meet the requirements, re-extract or purify. Next, enrich mRNA with Oligo(dT) beads, fragment the mRNA, synthesize first-strand cDNA with random hexamers, synthesize second-strand cDNA and purify the double-stranded cDNA. After that, perform end repair, add A-tail and ligate adapters on the purified double-stranded cDNA, select fragment size, PCR amplify and purify. Finally, inspect the library and perform sequencing with Illumina Novaseq 6000, convert the data and store it in FASTQ format.