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POSTPONED: Profiling the mammalian epigenome at single-cell resolution

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Friday, July 15, 2022
10:00 am - 11:00 am
Daniel Bartlett, PhD
COOPT Postdoc Candidate Seminar

Chromatin profiling in single cells has been extremely challenging and almost exclusively limited to histone proteins. In cases where single cell methods have shown promise, many require highly specialized equipment or cell type specific protocols and are relatively low throughput. Here, we combine the advantages of tagmentation, linear amplification and combinatorial indexing to produce a high throughput single cell DNA binding site mapping method that is simple, inexpensive, and capable of multiplexing several independent samples per experiment. Targeted Insertion of Promoters (TIP-seq) uses Tn5 fused to protein A to insert a T7 RNA polymerase promoter adjacent to a chromatin-bound protein of interest. Linear amplification of flanking DNA with T7 polymerase provides much higher starting material prior to sequencing library preparation and yields >10-fold higher unique reads per single cell compared to other methods.