Standard whole-genome library preparation works best with intact, high-molecular-weight DNA. Use 100 ng to 1 ug of input, with a 260/280 ratio near 1.8. Tagmentation-based kits can work with as little as 1 ng. Degraded DNA from FFPE or aged samples reduces library complexity. Check the fragment size distribution by gel or capillary electrophoresis before you start the prep.
NGS - Next Generation Sequencing
DNA Library Preparation Kits
DNA library preparation kits for short-read and long-read sequencing at any throughput
Pick the kit that matches your throughput, input, and indexing, and the rest of the run falls into place.
Library prep sets up everything downstream, so the choice should be an easy one. For whole-genome sequencing at scale, high-throughput 96-well plate kits give you multiple index sets for large-batch multiplexing, across human, plant, and animal genomes.
For lower-plex or more flexible work, compact kits with unique dual-index primers are a quick way in. Long-read multiplexing kits are ready when you need extended read lengths.
Tagmentation kits fragment and tag DNA in one step with an adapter-loaded transposase, so you skip separate fragmentation. Custom tagmentation reagents let you set your own adapter sequences, and spike-in indexed controls let you check run quality as you go. These formats run on Illumina short-read and PacBio long-read instruments.
Applications
Plasmid, amplicon or synthetic construct sequencing
For a fast, automation-ready run, reach for ExpressPlex 2.0. Its one-step TnX transposase workflow reduces bias and evens out coverage, with autonormalisation across the 96-well plate.
High-throughput human, plant and animal low-pass WGS and skim sequencing
For population-scale low-pass whole-genome or skim sequencing, AgriPrep keeps it simple. The 96-well plate kit handles human, plant, and animal samples in one run.
Whole-genome libraries where coverage uniformity matters
When you want high-complexity libraries with low duplication, the MosaiX kit gets you there in about 90 minutes. Directional TnX tagmentation suits whole-genome and targeted sequencing.
Long-Read Multiplexing
For PacBio long-read runs, the LongPlex Multiplexing Kit barcodes and pools your samples. It preserves fragment length for structural-variant calling and phasing.
Cell-free DNA and liquid-biopsy sequencing
For circulating tumour DNA and cell-free fetal DNA work, you build cfDNA libraries with the NEXTFLEX Cell Free DNA-Seq Kit from as little as 10 ng of cell-free DNA, in about three hours.
Custom adapter-loaded transposase
When your protocol needs specific adapter-loaded transposase sequences, Tagify reagents give you the control. Custom TnX and Tn5 sets and i5 UMI adapters are available.
Frequently asked questions
Which platform will my libraries run on?
The kits target Illumina short-read and PacBio long-read instruments, and some formats also run on Element. Match the kit to your sequencer before you start.
How much input DNA do I need?
Input depends on the kit. Tagmentation and cfDNA formats work from low nanogram amounts, for example the cfDNA kit runs from as little as 10 ng, while plate kits suit higher-throughput standard inputs.
What is the difference between ligation and tagmentation library prep?
Ligation attaches adapters in a separate step after fragmentation. Tagmentation fragments and tags DNA in one step with a transposase, which shortens the workflow and lowers the input needed.
Can I multiplex many samples in one run?
Yes. Plate kits offer multiple index sets for large-batch multiplexing, and unique dual-index primers reduce index hopping on patterned flow cells.
Unique dual indexes (UDI) give each sample a distinct combination of i5 and i7 adapter sequences. Both indexes must match for a read to be demultiplexed to the correct sample. This removes index-hopping artefacts that can occur on patterned flow cells. It matters most when you multiplex samples with very different read depths on the same run.
Tagmentation uses a transposase to fragment DNA and insert sequencing adapters in one enzymatic step. It is faster and needs less input material than traditional end-repair, A-tailing and adapter ligation workflows. The tradeoff is a fixed insertion bias pattern. For applications that need uniform coverage, ligation-based prep may work better.
A spike-in control is an indexed library of known sequence and quantity. You add it to the pooled sample at a defined dilution before loading. After sequencing, you check the fraction of reads that map to the spike-in sequence. This tells you the run efficiency, cluster density and demultiplexing accuracy, independently of sample quality variation.
