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What are the major steps in library construction with the Lotus DNA Library Prep Kit?
The major steps in library construction with the Lotus DNA Library Prep Kit are:
Enzymatic preparation. Fragmentation, end-repair, and dA-tailing of double-stranded DNA (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. For more details about amplification, refer to the Lotus DNA Prep Kit protocol available at www.idtdna.com/protocols.