Lotus DNA Library Prep Kit

One kit. Flexible workflow. Endless applications.

The Lotus DNA Library Prep Kit enables streamlined preparation of high-quality next generation sequencing (NGS) libraries from double-stranded DNA (dsDNA). The kit uses enzymatic fragmentation to generate libraries suitable for PCR-free, PCR-amplified, and targeted sequencing applications on Illumina platforms.

Get the uniform sample coverage you need without relying on expensive equipment
Regain valuable time with a fast, simple workflow
Create application-specific NGS libraries by adding IDT adapters and xGen products for target capture

The Lotus kit can be customized for your applications when combined with one of the many IDT adapter options (see Ordering section). Additionally, use of xGen hybridization capture products provides a complete NGS solution that takes you from sample preparation to sequencing.

Lotus™ DNA Library Prep Kit

Rapid, easy, and customizable enzymatic library preparation workflow for double-stranded DNA samples.

	Product	Catalog #	Sequence	Unit Size		Order Link
	Lotus™ DNA Library Prep Kit, 16 rxn	10001073				Click here to order
	Lotus™ DNA Library Prep Kit, 96 rxn	10001074

Adapters

To complete your libraries, you will need adapters. Use our Custom NGS Adapters and step-by-step configurator tool to create custom solutions that fit your exact application, or use preconfigured xGen Dual Index UMI Adapters—Tech Access, which are optimized for Illumina instruments and have the ability to run in 3 different schemes: single index, unique dual index, and unique molecular identifier modes.

Applications

The Lotus DNA Library Prep Kit produces high-quality next generation sequencing (NGS) libraries that are suitable for many applications including:

Whole genome sequencing (WGS)
PCR-free sequencing
Detection of germline inherited single nucleotide variations (SNVs) and insertions/deletions (indels)
Hybridization capture of target sequences (e.g., the exome or transcripts of interest)
Low frequency somatic variation detection of SNVs and indels
RNA-Seq starting with full-length, double-stranded cDNA input
Metagenomic sequencing

Method

This kit involves minimal enzymatic incubations and bead-based purification steps, thereby reducing sample handling and overall library preparation time to under 2 hours. The major steps are depicted in Figure 1 and are summarized as follows:

Enzymatic preparation. Fragmentation, end-repair, and dA-tailing of dsDNA are all performed in a single reaction.
Ligation of either full-length or stubby P5 and P7 adapters. When using full-length adapters, the final PCR step is optional and can be used to increase library yield. However, stubby adapters (sometimes called truncated adapters) require amplification with primers to incorporate sample indexing sequences and to add the P5 and P7 sequences.
Optional PCR amplification. Choose to amplify your libraries based on adapter and DNA input used. A bead-based cleanup removes oligonucleotides and small fragments after PCR.

Figure 1. Overview of Lotus DNA library preparation. Our easy enzymatic method takes you from sample to sequencing while eliminating the need for acoustic shearing methods that require instrumentation and extra time.

Kit overview

The Lotus kit comes with all the buffers, reagents, and enzymes needed for fragmentation, adapter ligation, and an optional PCR (to add index sequences, depending on adapter type, or to increase the amount of DNA for sequencing). You supply the DNA and add adapters that fit your goals. Table 1 provides additional specifications for using this kit.

Table 1. Specifications of the Lotus DNA Library Prep Kit.

Feature	Details
Kit sizes	16 reactions 96 reactions
Sample types (not provided)	Double-stranded DNA from fresh frozen tissue Genomic DNA Full-length double-stranded cDNA
DNA input range	1–250 ng
Suggested DNA insert size	Whole genome applications: 350 bp Hybridization capture: 200 bp
TA-ligation adapters (see Table 2)	Full-length: PCR amplification is optional Stubby (truncated): PCR is required to add index sequences

Adapters

The Lotus kit does not include adapters, giving you the flexibility to design and use your own adapter-indexing strategies. We offer many high quality, custom designed adapters and index primers that are application specific and ideal for use with the Lotus kit (Table 2).

Table 2. How to choose IDT adapters for common applications when using the Lotus DNA Library Prep Kit.

Application	Best choice	Alternatives
Whole genome sequencing, metagenomics, PCR-free sequencing (≥100 ng input)	TruSeq™-Compatible Full-length Adapters	xGen Dual Index UMI Adapters—Tech Access
Whole genome sequencing, metagenomics with PCR (1–250 ng input), exome sequencing, and targeted germline sequencing (SNVs, indels)	TruSeq™-Compatible Stubby Adapters and Indexing Primers	TruSeq™-Compatible Full-length Adapters or xGen Dual Index UMI Adapters—Tech Access
Low-level mutation detection, down to ~1% frequency	xGen Dual Index UMI Adapters—Tech Access	—
RNA-seq starting with full-length, double stranded cDNA input	xGen Dual Index UMI Adapters—Tech Access	TruSeq™-Compatible Full-length Adapters

Complete workflow for hybridization capture experiments

Lotus DNA Library Prep Kit paired with adapters from IDT create libraries that are compatible with xGen hybridization capture probes and reagents for when you only need to perform targeted sequencing (Figure 2). Here is a list of the key components you will need:

xGen Hybridization and Wash Kit
xGen Universal Blockers—TS Mix or 10 bp TS Mix
xGen Lockdown Panels and Probe Pools
xGen Gene Capture Pools
xGen Library Amplification Primers

Figure 2. Overview of hybridization capture sequencing workflow.

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Lotus DNA Library Prep Kit

Lotus™ DNA Library Prep Kit

Rapid, easy, and customizable enzymatic library preparation workflow for double-stranded DNA samples.

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Adapters

Applications

Method

Kit overview

Adapters

Complete workflow for hybridization capture experiments