The right answers to frequently asked questions
Find the answers to all our products and services by clicking the links below.
General Questions regarding Sample Submission, Ordering & Sample Shipment & Results can be found here:
Sample Preparation – for Amplicon sequencing on Illumina (short read). What are the sample submission guidelines?
Submit at least 25 µL purified PCR products sized 150-270 bp, with a concentration > 4ng/µL with a purity (A260/280) of 1.8-2.0. Amplicons must not contain TruSeq adapter sequence.
Submit at least 25 µL purified PCR products sized 280-570 bp, with a concentration > 1ng/µL with a purity (A260/280) of 1.8-2.0. Amplicons have to contain a TruSeq adapter sequence.
Samples can be in RNase-, DNase- and protease-free water, EB, or low TE buffer.
Please also refer to “2. Ordering & Sample Shipment” to read more about organizational topics on how your samples need to be prepared before being shipped to our sites
How should I purify my amplicons?
We recommend purifying your amplicons with commercially available kits based on DNA-binding beads or columns or enzymatic cleanup. DO NOT ship any primers with your samples or mixed into your samples
What types of samples can be submitted for amplicon sequencing?
Submit purified PCR products with a size range of 150 – 270 bp (NovaSeq) or 280 – 570 bp (MiSeq). The amplicons should ideally produce a single band on a gel for optimal results.
Can I submit amplicons outside the respective size ranges?
No, only amplicons within our size ranges 150 – 270 bp (NovaSeq) or 280 – 570 bp (MiSeq)) are accepted. For larger products, please consider our ONT amplicon sequencing services. If you are unsure about product selection, please reach out to your sales representative.
How can I multiplex multiple amplicons within a single sample?
For 150-270 bp amplicons sequenced on our NovaSeq, please use our Inline Tag Design Tool : NGS Adapter Ligation Oligos (eurofinsgenomics.eu). When ordering the service, select the optional service “Tag sorting – Eurofins Genomics Design”. You will receive sorted sequences (FASTQ).
- 192 individual inline tags are available, each consisting of 10 nucleotides added to the forward primer.
- There is a minimum edit distance of 4 to prevent misassignments due to sequencing errors.
- The tags are optimized for both low and high sample pools.
- All NGSgrade Oligos undergo specific processes to ensure cross-contamination-free and high-purity performance.
- For multiplexing 24 samples with the same target sequence, 24 forward primers (each with a unique inline tag) and one reverse primer (without tags) are required.
What are TruSeq adapter sequences?
The adapter sequences are:
Forward: 5’-ACACTCTTTCCCTACACGACGCTCTTCCGATCT-3’
Reverse: 5’-GACTGGAGTTCAGACGTGTGCTCTTCCGATCT-3’
I have mixed amplicons in a single sample that I want to sequence. Can I get the unique sequences from this mixture, and how can your service help with this?
Yes, you can send in a single sample containing mixed amplicons for sequencing. Our Unique Sequence Analysis service is specifically designed to handle such samples. By using advanced sequencing and bioinformatics techniques, we can accurately separate and identify each unique sequence, ensuring you get comprehensive and reliable results from your mixed amplicon sample.