What should I do if my library yields are low (less than 0.5 nM)?

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Low library yields are unavoidable in some experiments (e.g. mapping a low abundance target and/or using low cell numbers). For low-concentration libraries, consider the following options:

  • Concentrate library for sequencing: If library prep cannot be repeated, use a Speedvac to increase the library concentration and add as much of the library as possible to the sequencing pool. Deeper sequencing is recommended to ensure sufficient read depth for the the low-concentration library.

  • Repeat library prep: Use more CUT&RUN-enriched DNA input to improve yields for sequencing. As a last option, you can also consider increasing your PCR cycle number—for more on why this is discouraged, please see this article. Note that increasing the number of PCR cycles may lead to higher read duplication rates and require deeper sequencing to capture read diversity.

In both cases, deeper sequencing is strongly recommended.