2025-09-09
ENA first public
ERC000011
ENA-CHECKLIST
Duke-NUS Medical School
INSDC center name
public
INSDC status
ERS20852602
SRA accession
E-MTAB-14343:GENTYF-B20-D0
Submitter Id
42
year
age
ArrayExpress
broker name
Seropositive
clinical history
not collected
collection date
human
common name
Protocols: Blood was collected in Tempus tubes from subjects, and Tempus tubes were stored at -80deg until RNA extraction. We performed gene expression analysis of dengue seropositive versus dengue seronegative subjects. RNA extraction from whole blood was performed using the Tempus Spin RNA Isolation kit (Thermo Scientific). RNA was eluted in 90ul of RNase-free water. 1 μg total RNA was used for following library preparation. The poly(A) mRNA isolation was performed using Oligo(dT) beads. The mRNA fragmentation was performed using divalent cations and high temperature. Priming was performed using Random Primers. First strand cDNA and the second-strand cDNA were synthesized. The purified double-stranded cDNA was treated to repair both ends and add a dA-tailing in one reaction, followed by a T-A ligation to add adaptors to both ends. Size selection of Adaptor-ligated DNA was then performed using DNA Clean Beads. Each sample was then amplified by PCR using P5 and P7 primers and the PCR products were validated.
description
adult
developmental stage
normal
disease
not collected
geographic location (country and/or sea)
GENTYF-B20
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
Homo sapiens
scientific_name
male
sex
GENTYF-B20-D0
title