GSM5333714: S30 Glomeruli: Saline + Control; Mus musculus; RNA-Seq
Mice were sacrificed by cardiac perfusion. Kidneys were extracted and digested by GentleMACS. Digested kidneys were further treated by collagenase and hyaluronidase and passed through cell strainers. The glomeruli were collected and lysed in lysis buffer using an RNA isolation kit (Zymoresearch). The second strand cDNA synthesis was performed using DNA Polymerase I and RNase H. The synthesized cDNA was fragmented randomly and the fragments are purified and resolved with EB buffer for end repair and single nucleotide A (adenine) addition. Thereafter, the short fragments were connected with sequencing adapters. The double-stranded cDNA library is completed through size selection and PCR enrichment.