title: GSM7247699: SQLE_J82 repeat 3; Homo sapiens; RNA-Seq
library_construction_protocol: Total RNA was extracted from the tissue using TRIzol® Reagent according the manufacturer's instructions (Invitrogen) and genomic DNA was removed using DNase I (TaKara). RNA degradation and contamination was monitored on 1% agarose gels. Then RNA quality was determined by 2100 Bioanalyser (Agilent Technologies) and quantified using the ND-2000 (NanoDropTechnologies).   (TaKara). RNA degradation and contamination was monitored on 1% agarose gels. Then RNA quality was  determined by 2100 Bioanalyser (Agilent Technologies) and quantified using the ND-2000 (NanoDrop  Technologies). Illumina Novaseq 6000 platform was used to randomly fragment mRNA and isolate small fragments of about 300 bp by magnetic bead screening. Under the action of reverse transcriptase, random hexamers were added to synthesize one-strand cDNA using mRNA as template, followed by two-strand synthesis to form a stable double-strand structure. The cDNA structure of the double strand was A sticky End, which was made into A flat End by adding End Repair Mix, and then an "A" base was added to the 3 'End for joining the Y-shaped connector. The library was enriched and 15 cycles were amplified by PCR. Then, recover the target band by 2% agarose gel, quantify by TBS380 (Picogreen), and mix on the machine according to the proportion of data. Bridge PCR amplification was performed on cBot to generate clusters. Sequencing was performed by Illumina HiSeq xten platform (PE library, read length 2×150bp).