Custom Metagenome Amplification Sequencing

Preparation of custom metagenomic amplicon libraries targeting diverse population of pathogens  of any specific gene, Archaea, Cyanobacteria and internal transcribed spacer (ITS) regions for studying phylogeny and taxonomy from environmental samples.

Methodology

Sample Requirement:

• Submit 1-3 µg of RNA-free meta genomic DNA (NGS grade) of high molecular weight,  Nanodrop A_260/280 ratio >1.8; A_260/230=2.0-2.2; and at a concentration of ~50 ng/µl
• Or soil, plant/ human microbiome, water or sludge samples, shipped in cool packs to Eurofins facility.

Sample QC:

• RNA free NGS grade gDNA isolation will be carried out using commercially available DNA isolation kit.
• Quality check will be carried out using agarose electrophoresis and NanoDrop. Quantification will be carried out using Qubit 3.0 Fluorometer.

Library Preparation:

• Amplicon optimization will be carried out before sequencing library preparation.
• Sequencing ready customized amplicon library will be prepared and indexed using Nextera XT index kit of illumina.
• Library validation will be carried out using Agilent 4200 Tape station kit.

Sequencing:

Sequencing will be carried out on Illumina MiSeq platform using 2 x 300 bp paired end sequencing chemistry.

Deliverable

• Raw data will be available for download as a compressed archive of FASTQ files for each sample.
• Comprehensive compiled report and data set will be shared in link or Pen Drive or Hard Disk.
• For analysis please visit Bioinformatics Services

Turnaround time

5-7 weeks after arrival of your samples and all necessary information. It depends upon data size, scope, technology selected, number of samples and complexity of the project. The TAT is offered expecting no biological or technical difficulties for processing of all project samples.