Ranger® Technology offers turnkey automated size selection, and can be seamlessly leveraged to provide high throughput fragment length analysis and fluorescence assays. A selection of applications that have already benefited from Ranger® Technology are discussed below
Bioanalyzer 2100 (Agilent) electropherogram of proxy sample that was size selected using Ranger® Technology
Cell Free DNA (cfDNA) – Liquid Biopsy and NIPT
Sequencing of cfDNA is an area of intense interest to the DNA sequencing community. Characterisation of cfDNA from non-normal sources can provide diagnostic insights for pregnancies (NIPT), cancer (liquid biopsy), and many other clinical and research scenarios.
New research has shown that the cost of diagnosing genetic disorders and cancer through sequencing can be improved through automated size selection. Size selection looks to recover cfDNA from non-normal tissues while excluding that from healthy, normal cells.
References
[citation #1: Underhill et al] [citation #2: Mouliere et al].
Ranger® Technology allows you to automate the process of fetal DNA enrichment through size selection, and has been show to secure a recovery yield averaging over 70% in various NIPT applications
Yourgene Health works with academic and clinical collaborators to realis e the benefits of size selecting for cfDNA applications. At the AACR Virtual Annual Meeting in 2020, the LabCorp Speciality Testing Group presented their work demonstrating that size selection of cfDNA using Ranger® Technology increases the proportion of tumour-specific variants in cancer patients. You can listen to Dr Kimberly Holden from LabCorp talk about this work in our on-demand webinar.
Our latest collaboration with Dr. Sabine Hellwig, Dr. Hunter Underhill and team at the University of Utah demonstrated that automated size selection using Ranger® Technology was able to enrich for mutant allele fractions , while concomitantly reducing the overall false-positive rate. This work was published in 2018 in PLOS ONE, and demonstrates how Ranger® Technology can be used to automate the process of cfDNA size selection to better outcomes for cancer patients.
Learn more about the work of Dr Underhill and group in this white paper, which explores the current state and future directions of liquid biopsy. You can also read more from Dr Underhill’s work comparing the performance of Ranger® Technology with traditional PAGE extraction here.
Metagenomics
Metagenomic sequencing of bacterial communities can involve so-called ‘tagmentation’ approaches to library preparation. This process relies on the tight control of an enzymatic process that fragments larger DNA fragments into smaller pieces, the length of which cluster around a desired size.
Often, unavoidable deviations can lead to inconsistent fragment length profiles between different samples. This in turn can produce sequencing reads that are undesirably short, greatly limiting the data output of the sequencing run. Dr. Miguel Uyuguari-Diaz has leveraged Ranger® Technology to improve data output for his metagenomic research on antibiotic resistance genes
References
[citation #1: Uyaguari-Diaz et al] [citation #2: Uyaguari-Diaz et al].
Small Nucleic Acid Work
The operating principle behind Ranger® Technology was developed with the intent of recovering small nucleic acids whilst confidently rejecting adaptermers that were close in size to the target. Work with microRNA-SEQ experiments greatly benefits from size selection, which can be accurately conducted at any scale with Ranger® Technology.
Bioanalyzer 2100 (Agilent) electropherogram of proxy sample that was size selected using Ranger® Technology
Bioanalyzer 2100 electropherogram of proxy sample post size selection with Ranger® Technology
Overlay of Bioanalyzer 2100 electropoerograms of proxy sample input (pre size selection) and output (post size selection with Ranger® Technology
Large Fragment Isolation
The growing interest in sequencing of large DNA fragments has led to increased throughput requirements for selective exclusion of smaller DNA fragments that can cause problems for the application. Any life science application using large DNA fragments (typically defined as >5kbp in length) can benefit from quality control measures, such as fragment length analysis. Ranger® Technology can process up to 96 samples in parallel for exclusive recovery of large DNA fragments via analysis and subsequent size selection. Size selections that reject fragments below a specific cut-off threshold can be conducted for samples up to 4 ug in total mass.
You can read more about how Ranger® Technology has been shown to improve data yield from Illumina Nextera XT Library Preparation kits by definitive exclusion of short fragments here.
DNA Quality Control
Ranger® Technology comes with the hardware complement that is necessary for it to act as a DNA quality control instrument. In addition to being compatible with Yourgene reagent kits that enable high throughput fragment length analysis, the system can be used as a plate reader for the evaluation of solution-based fluorometric assays for the quantification of DNA. Data from both assays are combined with Ranger® Technology software to improve upon the accuracy of concentration estimations from smear analyses. This in turn can improve balancing of libraries prior to multiplexed next generation sequencing.
Ranger® Technology is compatible with numerous commercially available fluorophores. Please contact us to discuss what viable options are.
Click here to see how the LightBench® can be used to evaluate solution-based fluorometric assays, offering unparalleled flexibility and performance against existing methodologies.