What if my yield of CUT&Tag DNA is too low to see on an Agilent Bioanalyzer or TapeStation system?

Due to the expected and typically low yield of CUT&Tag DNA, we recommend using all 30 µL of the CUT&Tag DNA sample (from Section VI of the CUT&Tag protocol #77552) for library amplification. While CUT&Tag DNA libraries generated for histone modifications typically show some signal on the Agilent Bioanalyzer or TapeStation systems, libraries generated for non-histone proteins such as transcription factors and cofactors often have very weak or even no visible signal on Bioanalyzer or TapeStation systems, but they still generate NGS results with high-mapping rates, high numbers of identified binding peaks, and decent signal-to-noise ratios across the whole genome. For CUT&Tag libraries where the Bioanalyzer or TapeStation system is unable to identify the average size of the library, we suggest using a size of 900 bp to intentionally pool more low-yield libraries than normal-yield libraries.

Last updated: September 12, 2024