P16_09
Protocols: RNA was isolated from temporal cortex tissue of 10 FTLD brains. 50 mg of flash-frozen tissue was homogenised in 700 µl of Qiazol (Qiagen) using a TissueRuptor II (Qiagen). Chloroform was added and RNA subsequently extracted following the spin-column protocol from the miRNeasy kit with DNase digestion (Qiagen). RNA was eluted off the column in 50 µl of RNAse-free water. Purified RNA was reverse transcribed with Superscript IV (Thermo Fisher Scientific) using either sequence specific primers containing sample-specific barcodes or random hexamers, following the manufacturer recommendations. Unique molecular identifiers (UMIs) and part of the P5 Illumina sequence were added either during first- or second-strand-synthesis (with Phusion HF 2x Master Mix) respectively. Three rounds of nested PCR using Phusion HF 2x Master Mix (New England Biolabs) were used to obtain highly specific amplicons for the UNC13A cryptic, followed by gel extraction and a final round of PCR in which the full length P3/P5 Illumina sequences were added.
ENA first public
2021-07-20
ENA last update
2021-07-20
External Id
SAMEA8268183
INSDC center alias
UNIVERSITY COLLEGE LONDON
INSDC center name
UNIVERSITY COLLEGE LONDON
INSDC first public
2021-07-20T00:14:55Z
INSDC last update
2021-07-20T00:14:55Z
INSDC status
public
Submitter Id
E-MTAB-10237:P16_09
age
68
broker name
ArrayExpress
common name
human
developmental stage
adult
disease
FTLD-TDPA
organism part
temporal cortex
sample name
E-MTAB-10237:P16_09
sex
male