GSM8772913: aNMP-2a0_1,Day7; Mus musculus; RNA-Seq
The heart was removed and frozen in liquid nitrogen.The tissue sample were homogenized the lysated using a rotor-stator homogenizer in Trizol. After DNA removal by AllPrep DNA spin column,the RNA were collected using a RNeasy spin column. After two washes, the RNA was dissolved in RNase-free water. After QC step, a certain amount of RNA samples are denatured at suitable temperature to open their secondary structure, and mRNA is enriched by oligo (dT) -attached magnetic beads. Then the RNAs were fragmented and cDNA were synthesized . Double-stranded cDNA fragments were subjected to end-repair, add A and adaptor ligation.The PCR reaction system and program are configured and set up to amplify the product. After Library QC step, single-stranded PCR products are produced via denaturation. Single-stranded cyclized products are produced, while uncyclized linear DNA molecules are digested. Single-stranded circle DNA molecules are replicated via rolling cycle amplification, and a DNA nanoball (DNB) which contain multiple copies of DNA is generated. Sufficient quality DNBs are then loaded into patterned nanoarrays using high-intensity DNA nanochip technique and sequenced through combinatorial Probe-Anchor Synthesis (cPAS).