T3
human gut metagenome
Protocols: Tissue specimens were collected from surgically resected tumor samples of patients, snap-frozen at -80°C, and transferred in liquid nitrogen. This study was approved by the Ethics Committee. Total RNA was extracted from the samples using the [Insert Kit Name, e.g., TRIzol reagent / Qiagen RNeasy Mini Kit] according to the manufacturer's instructions. The concentration and purity of the RNA were determined by measuring the absorbance at 260 nm and 280 nm using a NanoDrop spectrophotometer (A260/A280 ratio). RNA integrity was assessed by agarose gel electrophoresis to verify the presence of intact 18S and 28S ribosomal RNA bands. 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 then 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.
ENA first public
2026-01-01
INSDC center name
Hangzhou Hospital of Traditional Chinese Medicine, affiliated to Zhejiang University of Traditional Chinese Medicine
INSDC status
public
Submitter Id
E-MTAB-16416:T3
age
74
broker name
ArrayExpress
collection date
not collected
developmental stage
adult
disease
colon carcinoma
genotype
wild type genotype
geographic location (country and/or sea)
not collected
individual
Patient_03
isolate
not applicable
organism part
colon
sample name
E-MTAB-16416:T3
scientific_name
human gut metagenome
sex
female