Homo sapiens
Protocols: Lymphoblast cell lines were established with EBV immortalization for three affected individuals (1-III-16, 1-IV-9, and 1-IV-11) and one healthy spouse (1-III-24) from Family 1, three affected individuals (5-III-2, 5-III-3 and 5-IV-4) from Family 5 and three healthy controls (Ctrl1-3) Total RNA including small RNAs were extracted from immortalized lymphoblastic cells from each individual using the AllPrepR DNA/RNA/miRNA kit (Qiagen). Small RNA libraries were generated from 2 μg of total RNA using TruSeq Small RNA Sample Prep Kit (RS-200-0012/0024). The protocol uses RNA molecules that have a 5'-Phosphate and a 3ʹ-hydroxyl group. Briefly, in the first step, RNA adapters were sequentially ligated to each end of the RNA, first the 3ʹ RNA adapter that is specifically modified to target microRNAs and other small RNAs, then the 5ʹ RNA adapter. Small RNA ligated with 3ʹ and 5ʹ adapters were reverse transcribed and PCR amplified (30 s at 98°C; [10 s at 98°C, 30 s at 60°C, 15 s at 72°C] x 13 cycles; 10 min at 72°C) to create cDNA constructs. This process selectively enriched the fragments with adapter molecules on both ends. The last step was an acrylamide gel purification of 140-145 nt amplified cDNA constructs (corresponding to cDNA inserts + 120 nt adapters). Final libraries were checked for quality and quantified using capillary electrophoresis.
ENA-FIRST-PUBLIC
2019-09-16T04:03:09Z
ENA-LAST-UPDATE
2019-09-04T14:00:34Z
External Id
SAMEA5933685
INSDC center name
IGBMC, GenomEast plateform
INSDC first public
2019-09-16T04:03:09Z
INSDC last update
2019-09-04T14:00:34Z
INSDC status
public
Submitter Id
E-MTAB-8300:CSDN20
broker name
ArrayExpress
cell line
EBV-immortalized lymphoblast cell line
cell type
lymphoblast
common name
human
disease
normal
individual
CSDN20
organism part
lymphoid tissue
sample name
E-MTAB-8300:CSDN20
scientific_name
Homo sapiens