Experts in amplicon sequencing

Characterise all your sequence variants with our amplicon sequencing service

By taking advantage of high-throughput sequencing we can sequence all the gene variants of heterogeneous multi-copy regions. With our amplicon sequencing service you will be able to recover all the diversity regardless of the number of indels or point mutations. This is extremely useful when studying gene families such as the vertebrate major histocompatibility complex (MHC) or the ribosomal non-transcribed spacers (ITS1 and ITS2). To do so we design primer pairs at the flanking regions of the genomic region under study. Then, we prepare amplicon libraries which are sequenced using the Illumina MiSeq technology. Finally, using high-throughput methods, we analyse the sequencing results. Please note that currently, the size of the genomic regions should not exceed 550 base pairs.

How we work

Amplicon sequencing workflow

Step 1: We isolate DNA from your samples. We have adapted different DNA isolation protocols, depending on the starting biological material.

Step 2: We prepare amplicon libraries for the target genomic region(s). For this, we use universal or in-house-developed primer pairs.

Step 3: We sequence the amplicon libraries using the Illumina MiSeq technology.

Step 4: We analyse the data obtained in the previous step to obtain all the variants occurring in your samples.

For your convenience, we can carry out the entire project or only those steps you require. As a norm, steps 2 and 3 should be ordered jointly. Please contact us for further information and pricing.

What you receive

A report with a summary of the methods followed.

The raw high-throughput sequencing files.

The results of the bioinformatic analysis including demultiplexing, pre-processing and quality control of the raw data, chimaera removal, clustering of the reads into variants, number of reads obtained per variant, and the generation of rarefaction curves.

If you require additional analyses, please let us know and we will do our best to meet your needs.

At no additional cost, our project specialists will be available to you after project delivery and during the editorial process of your scientific articles. We offer up to five 60-min sessions per project, depending on the project size.

What we need

We can start working by using either your tissue samples, or DNA which has already been extracted.

If you are sending tissue samples, please make sure that they have been properly preserved for DNA isolation (ethanol, silica gel, frozen, or another suitable preservation method). If required, we can provide sampling kits and sample collection guidelines to ensure that your samples arrive at our lab in optimal conditions.

If your DNA has already been extracted, we ideally require 20 μL of DNA in TE buffer or water at a minimum concentration of 10 ng/μL, and 260/280 and 260/230 absorbance ratios higher than 1.8.

Please note that, depending on the specifics of your project, these requirements may vary. Please contact us for the specific requirements of your project.

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