We recently introduced Cleavage Under Targets & Tagmentation (CUT&Tag), an epigenomic profiling strategy in which antibodies are bound to chromatin proteins in situ in permeabilized nuclei. These antibodies are then used to tether the cut-and-paste transposase Tn5. Activation of the transposase simultaneously cleaves DNA and adds adapters (‘tagmentation’) for paired-end DNA sequencing. Here, we introduce a streamlined CUT&Tag protocol that suppresses DNA accessibility artefacts to ensure high-fidelity mapping of the antibody-targeted protein and improves the signal-to-noise ratio over current chromatin profiling methods. Streamlined CUT&Tag can be performed in a single PCR tube, from cells to amplified libraries, providing low-cost genome-wide chromatin maps. By simplifying library preparation CUT&Tag requires less than a day at the bench, from live cells to sequencing-ready barcoded libraries. As a result of low background levels, barcoded and pooled CUT&Tag libraries can be sequenced for as little as $25 per sample. This enables routine genome-wide profiling of chromatin proteins and modifications and requires no special skills or equipment.

我们最近推出了标靶切割和标记 (CUT&Tag)，这是一种表观基因组分析策略，其中抗体与透化细胞核中的染色质蛋白原位结合。然后使用这些抗体来束缚剪切粘贴转座酶 Tn5。转座酶的激活同时切割 DNA 并添加接头（“标记”）以进行双端 DNA 测序。在这里，我们引入了一种简化的 CUT&Tag 方案，该方案可抑制 DNA 可及性伪影，以确保抗体靶向蛋白的高保真图谱，并提高当前染色质分析方法的信噪比。简化的 CUT&Tag 可以在单个 PCR 管中进行，从细胞到扩增文库，提供低成本的全基因组染色质图谱。通过简化文库制备，从活细胞到可测序的条形码文库，CUT&Tag 在工作台上花费的时间不到一天。由于背景水平较低，条形码化和混合的 CUT&Tag 文库的测序成本每个样品只需 25 美元。这使得染色质蛋白和修饰的常规全基因组分析成为可能，并且不需要特殊技能或设备。