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 Dual Index 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 Dual Index UMI Adapters–Tech Access||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 Dual Index UMI Adapters–Tech Access||More sensitive – use UMI consensus analysis to remove sequencing errors|
|RNA-seq starting with full-length,
double-stranded cDNA input||xGen Dual Index UMI Adapters–Tech Access||More quantitative – UMI is used to remove PCR duplicates|
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