title: GSM1194804: GLD-2; Homo sapiens; RNA-Seq
library_construction_protocol: RNA was purified using TRIzol Reagent (Invitrogen (Life Technologies), 15596-018) according to manufacter’s insctructions. Total RNA was quantified on the NanoDrop 2000c Spectrophotometer (Thermo Scientific) and their quality was evaluated on the RNA Nano Chips (Agilent 2100 bioanalyzer, Agilent, 5067-1511). libraries were prepared by FASTERIS. 1)Transcripts purification; 2) Transcripts breaking in presence of Zn; 3) Double-stranded cDNA synthesis using random primers and RNase H; 4) Ends repair; 5) 3’ A addition; 6) Ligation of adapters; 7) Gel purification to isolate fragments of specified inserts size; 8) PCR amplification to generate the DNA Colonies Template Library, library purification; 9) Library quantification and dilution at 10 nM. 10 )Library Quality Control, done by titration run on the HiSeq. All the libraries to be quality controlled are multiplexed in 1 channel of 1x50 bp run on the HiSeq. After demultiplexing of the reads according to the indexes, we typically provide 50 fasta reads and 100'000 fast-q reads from each library. We can then blast the fasta sequences to check the quality of the library. In addition, the titration run helps us for quantification of the libraries.
attributes.tag: GEO Accession
attributes.value: GSM1194804