AutoCUT&Tag: streamlined genome-wide profiling of chromatin proteins on a liquid handling robot v1
Résumé
The CUT&Tag method is based on antibody tethering of the Tn5 transposase to profile the genome-wide occupancy of DNA-binding proteins, histone modifications and chromatin modifying proteinsin situ starting from relatively low cell numbers (1-100K cells). Activation of tethered transposase integrates library adapters at the sites of bound chromatin factors. By taking advantage of protocol modifications that enable Tn5 release and PCR in the same sample reaction chamber, we adapted CUT&Tag to an automated 96 well format. We provide programs and equipment specifications for performing AutoCUT&Tag on a Beckman Coulter Biomek FX liquid handling robot equipped for magnetic separation and temperature control, but in principle the method could be easily translated to other liquid handling units. Using this automated protocol a single operator is able to generate up to 96 CUT&Tag libraries in a single day that are ready to be pooled and sequenced.
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