Preparation of amplicon libraries targeting any specific gene, Archaea, Cyanobacteria and internal transcribed spacer (ITS) regions for studying phylogeny and taxonomy from environmental samples.
- Submit 1-5 µg of RNA-free meta genomic DNA (NGS grade) of high molecular weight, Nanodrop A260/280 ratio >1.8; A260/230=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.
- RNA free NGS grade gDNA isolation will be carried out using commercially available DNA isolation kit.
- Further quality checking will be carried out using agarose electrophoresis and NanoDrop. Quantification will be carried out using Qubit 3.0 fluorometer.
- 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 High Sensitivity D1000 Screen tape using Agilent Tape station kit.
- Sequencing will be carried out on illumina Miseq using 2 x 300 bp paired end sequencing chemistry to generate required amount of data.
- Delivery of data up to ~5, 00,000 paired end reads per sample.
- Sequence data (base call files or bcl files) generated from the sequencer are de-multiplexed if required and converted to FASTQ files.
- Raw data will be available for download as a compressed archive of FASTQ files for each sample.
- Raw data can be post processed upon request.
- 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.