G‑SPLAT: Genomic SPLAT for Whole Genome Sequencing

G-SPLAT is a library preparation technique designed for low-quality or limited-input DNA, with applications in whole-genome sequencing, metagenomics, and and other challenging sequencing projects.

G‑SPLAT (Genomic SPLinted Ligation Adapter Tagging) is a streamlined whole genome sequencing (WGS) library preparation method developed by our R&D team at NGI Sweden. It is directly adapted from our BSK‑SPLAT protocol, which is used for whole genome bisulfite sequencing (WGBS), but in this version the bisulfite conversion step is omitted. By retaining the efficient adapter ligation strategy for ssDNA (or denatured dsDNA), G-SPLAT is optimized for a wide range of challenging DNA samples, including single-stranded, double-stranded, degraded, or low-input material. Standard DNA input is 10–200 ng; however, inputs as low as 1 ng are feasible for this method. Its performance is on par with commercial kits, but it is particularly advantageous for single-stranded DNA, where alternative methods often fail.

Key Features

• Same SPLAT ligation chemistry, adapted for genomic DNA
• Ideal for difficult samples, including for example: FFPE, metagenomic samples, eDNA, cell free DNA, ssDNA-viruses.
• Useful for fragmentomics as it preserves native ends
• Cost-efficient and reproducible like BSK‑SPLAT

Sample requirements

• Sample type: DNA
• Amount of sample: >200ng
• Volume: 25-50 µL
• Amount of material needed for preparing one library: 10-200ng (standard)

How to evaluate the sample quality

We check samples upon arrival; however, we strongly recommend that users perform their own quality control prior to shipment.
Checking the concentration
Please use fluorometric measurements (e.g. Qubit, Quant-it), not absorbance measurements (Nanodrop, spectrophotometer).

Checking the quality and purity
The DNA should be in water or EB buffer, of high quality, and ideally have an OD260/280 of 1.8-2.

If your samples are below the required thresholds, please get in touch.

What we do with your samples

Once your samples arrive at NGI, we start by performing a quality control (QC) step in which we make sure the samples meet our requirements.

If the samples fail this quality control step, we will contact you to discuss possible options. If you choose to proceed with samples that do not meet the acceptance criteria, this will be at your own risk. NGI will make only one attempt at library preparation, and the library preparation fee will be charged regardless of outcome.

Samples that pass quality control will proceed to the library preparation queue.

Library preparation

DNA is fragmented by Covaris, denatured, and ligated to adapters using a single-stranded DNA ligation strategy, followed by bead-based cleanup steps. Libraries are PCR amplified with unique dual indexes and purified to generate sequencing-ready libraries.

Library QC and sequencing

We evaluate the concentration of the library using qPCR and inform you of the QC status of each sample. Once the libraries have passed this QC step, they are normalised, pooled, and queued for sequencing.

The sequencing will be carried out following the setup stated in the agreement.

Expected results

G‑SPLAT is a cost efficient, in-house alternative for whole-genome sequencing (WGS) when working with challenging DNA, offering flexibility, high performance, and reliable results. For more information regarding this technique, please contact our project coordinators.

Last Updated: 28th January 2026