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
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.
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