Area

Sequencing


The ISS sequencing service is equipped with the Ion GeneStudio S5 System, a next-generation sequencing (NGS) system based on semiconductor technology (Ion S5 chip) that enables simple and fast multi-targeted sequencing workflows with high accuracy at reduced costs.

Whenever a single nucleotide is incorporated during DNA polymerization, a hydrogen ion is released: the resulting pH variation is detected by the instrument within each CHIP well. The nucleotide incorporation is associated to each pH variation and the chemical signal is then converted into digital signal. Since the growing complementary DNA strand is flooded with a single species of nucleotide at a time, a signal is detectable only when a nucleotide is successfully incorporated.

The Ion GeneStudio S5 System enables multiplexing of up to 96 fragment libraries on a single sequencing chip thank to the use of specific molecular barcodes for sample-labelling.

For DNA analysis, the required starting material is about 10-100 ng (at 10 ng/µl).

The availability of 4 chip sizes (Ion chip 510, 520, 530 and 540) fits with many different applications and throughput needs and allows to achieve from 2–80 million reads per run.


The instrument The semiconductor chip

On the left: the instrument – on the right: the semiconductor chip



NGS: mechanism of action

Il principio di funzionamento


Applications

Ion GeneStudio S5 technology is mainly indicated for the following applications:

  • Microbial sequencing (DNA, RNA , also de novo);
  • whole exome and transcriptome;
  • Targeted DNA and RNA sequencing based on the Ampliseq technology,
  • Small RNA and miRNA sequencing;


To ask for the NGS service:

  1. Preliminary contact the Sequencing area staff;
  2. fill out the online form 'Service Request' briefly illustrating the scientific problem, type of sample (DNA, RNA etc.), number of samples to be sequenced etc.;
  3. Plan a meeting with the sequencing area staff to discuss methods of analysis, costs, timing etc.


Contact: Fiorella Ciaffoni
+39 06 4990 3075