GSM1214554: MSUS 3; Mus musculus; RNA-Seq
RNA was extracted according to the standard Trizol protocol after purifying samples using elutriation centrifugation. The quantity and quality of RNAs were determined by Qubit® fluorometer (Life Technologies) and Agilent 2100 Bioanalyser (Agilent Technologies), respectively. Small RNA libraries were prepared according to a modified Illumina v1.5 protocol. Briefly, small RNAs of <50 nt were purified on an acrylamide gel. Universal miRNA cloning linker (New England Biolabs) instead of 3´ adapters and then 5’ Illumina adapters ( CTGTAGGCACCATCAAT)were single-stranded ligated with T4 truncated RNA and T4 ligase respectively. The constructs were purified on an acrylamide gel to remove empty adapters then reverse-transcribed and PCR-amplified. The primers used for cDNA synthesis and PCR were designed to insert an index in the 3- adapter. Multiplexing indexes were as follows: Sample 1: ACACCA, Sample 2: CTTTCT, Sample 3: GAGTTG, Sample 4: CAAGTA, Sample 5: GATGGC, Sample 6: GGCACG, Sample 7: CGACAG.
GEO
GSM1214554