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Hairpinloop adapters during library preparation

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  • Hairpinloop adapters during library preparation

    Hello,

    Can you please help me understand what is the function of NEBNext adapters (hairpinloop adapters) during library preparation. Please correct me if I am wrong, as much as I understand the sequencing adapters consists of 1. Sequencing primers (read 1 and read 2), 2. Unique indexes and 3. indexes complimentary to P5/ P7.

    What is the function of these NEBNext adapters or any adapters for that matter which we ligate separately from indexes? Are they essentially the sequencing primers?

    Also during library preparation, why do we perform adapter ligation and indexing separately? Can we not attach the complete adapter in one go?

    I am new to library preparation, I would really appreciate your help.


    Thank you very much.
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