title: GSM9240239: ZBTB20,cKO,B1; Mus musculus; RNA-Seq
library_construction_protocol: Using sterile instruments, the SVZ area from the mouse brain was extracted and then added to trizol. It was then stored in liquid nitrogen. Total RNA was used as input material for the RNA sample  preparations. Briefly, mRNA was purified from total RNA using poly-T  oligo-attached magnetic beads. Fragmentation was carried out using  divalent cations under elevated temperature in First Strand Synthesis Reaction Buffer(5X). First strand cDNA was synthesized using random  hexamer primer and M-MuLV Reverse Transcriptase(RNase H-). Second strand cDNA synthesis was subsequently performed using DNA Polymerase I and RNase H. Remaining overhangs were converted  into blunt ends via exonuclease/polymerase activities. After adenylation  of 3' ends of DNA fragments, Adaptor with hairpin loop structure were  ligated to prepare for hybridization. In order to select cDNA fragments  of preferentially 370~420 bp in length, the library fragments were  purified with AMPure XP system. Then PCR was performed with Phusion High-Fidelity DNA polymerase, Universal PCR primers and Index (X) Primer. At last, PCR products were purified (AMPure XP  system) and library quality was assessed on the Agilent Bioanalyzer 2100 system.