Frequently Asked Questions
1. What is contiguous amplicon sequencing?
Contiguous amplicon sequencing involves sequencing the
2. When does GENEWIZ need to fragment amplicons for sequencing and what does this impact?
GENEWIZ recommends fragmentation with amplicons >570bp. Amplicons are randomly fragmented, with NGS adapters ligated to each individual fragment. Once fragmented, phasing information is lost and library diversity studies are not possible. Studies are focused solely on variant frequency detection across the full reference sequence.
3. What is “PhiX” and why is it needed for amplicon sequencing?
PhiX is a well-studied bacteriophage that has balanced genomic content. Most amplicons have low diversity at the 5’ and 3’ ends, which inherently reduces Illumina data quality based on current
4. How much PhiX is typically spiked-in to a sequencing run and how does this affect my data?
We recommend spiking in ~10-30% PhiX, depending on the original library diversity. The PhiX sequences are removed from the data reported to the
We have created a bioinformatics pipeline that reports every unique sequence and their relative abundance at the molecular level. This is particularly effective for library diversity studies. Please ask our NGS team for an example report.
|Amplicon Length||Library Preparation Method||MiSeq Configuration Recommended|
6. How do I contact GENEWIZ for a technical consultation for my project?
GENEWIZ's NGS Team is composed of Ph.D. scientists who can help you optimize your project design and provide consultation to answer any questions. For any form questions that require support, please contact the NGS team at [email protected], toll-free at 877-436-3949 ext. 1, or directly at +1-908-222-0711 ext. 1.
7. Can GENEWIZ recommend the best data delivery option?
Yes, GENEWIZ will recommend the best data delivery option based on the platform and project details. Data delivery options include: