2024-10-20
ENA first public
ERC000011
ENA-CHECKLIST
Tianjin Chengjian university
INSDC center name
public
INSDC status
ERS21119589
SRA accession
E-MTAB-14496:Sample 1
Submitter Id
ArrayExpress
broker name
not collected
collection date
Protocols: Dehydrated sludge collected from a PWWTP in Guangzhou Take a 1.5 ml centrifuge tube, add 200 µL of pretreatment solution and three glass beads, then add the appropriate amount of bacterial sample and put it into the grinder and grind it until well done. Add 20 µL of Proteinase K solution, mix well, and leave at 37°C for 30-60 min. Add 200 µL of lysate, mix well inverted, and leave at 70°C for 10 min. Add 200 µL of anhydrous ethanol, mix well inverted, and centrifuge briefly to remove droplets from the inside of the cap. Pass through the adsorption column, wash once with washing solution and twice with rinsing solution. Allow the column to stand at room temperature for 5-10 minutes to thoroughly dry any rinse solution remaining in the adsorbent material. Transfer the adsorbent to a new centrifuge tube, add 50-100 µL of ddH2O dropwise to the centre of the adsorbent membrane, leave at room temperature for 5-10 min, centrifuge at 12,000 rpm for 2 min, and collect the solution in the centrifuge tube. Firstly, the DNA was screened by magnetic beads for large fragments; then, library construction was carried out: a) for each library, about 1ug of DNA was taken after magnetic bead screening for damage repair and end repair; b) the recovered repaired DNA was purified by the magnetic bead method, and the concentration was determined by Qubit and the recovery rate was calculated; c) each sample was added with the corresponding barcode, and the barcode ligation product was purified by magnetic beads; d) the barcode ligation product was purified for multiple samples; e) the barcode ligation product was purified by magnetic beads. d) Calculate the pooling ratio for multiple barcode ligations according to the data volume requirement, and pool them proportionally; e) Junction ligation, so that each DNA fragment carries a specific junction sequence that can be recognised; f) Purify the ligated sample DNA by magnetic bead method, and then determine the concentration by Qubit and calculate the recovery rate. Finally, sequencing was performed.
description
not collected
geographic location (country and/or sea)
not applicable
isolate
mixed sample
organism
mixed sample
organism
mixed sample
scientific_name
Ochrobactrum intermedium and Stenotrophomonas acidaminiphila
strain
Sample 1
title