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.
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.
The Lotus DNA Library Prep Kit produces high-quality next generation sequencing (NGS) libraries that are suitable for many applications including:
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:
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.
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.
|Kit sizes||16 reactions|
|Double-stranded DNA from fresh, frozen tissue |
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
Complete your library prep with our selection of ready-to-use xGen UDI-UMI Adapters or xGen Stubby Adapter and UDI Primer Pairs (Table 2). We also offer many high-quality, custom adapters and index primers that are application-specific and ideal for use with the Lotus kit.
Table 2. How to choose IDT adapters for common applications when using the Lotus DNA Library Prep Kit.
|Whole genome sequencing (WGS), metagenomics, PCR-free sequencing (≥100 ng input)||xGen UDI-UMI Adapters||Full-length adapters – required for PCR-free applications|
|WGS, metagenomics with PCR (1–250 ng input), exome sequencing, and targeted germline sequencing (SNVs, indels)||xGen Stubby Adapter and UDI Primer Pairs||Simple workflow – prepare your ligation master mix with a universal adapter and introduce unique dual index sample barcodes via PCR|
|Low-level mutation detection, down to ~1% frequency||xGen UDI-UMI Adapters||More sensitive – use UMI consensus analysis to remove sequencing errors|
|RNA-seq starting with full-length,
||xGen UDI-UMI Adapters||More quantitative – UMI is used to remove PCR duplicates|
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:
The Lotus DNA Library Prep Kit produces consistent genome coverage with low bias across samples (Figure 3).
IDT adapters and xGen Lockdown Probes and Panels are manufactured using stringent, proprietary methods that are critical for producing high-quality oligos for NGS applications. When these adapters and hybridization capture probes are used with the Lotus Kit for targeted sequencing, results show consistent, highly uniform, sequence coverage (Figures 4–5).
The observed representation of each genome was consistent across a range of inputs (1, 10, and 25 ng) and correlated well with expected results (Figure 6A). Consistent genome coverage was observed across samples with varying levels of GC content (Figure 6B). The variability in size of the genomes, input amount, and GC content do not influence results.