Embedded Files
●Select Regulatory Regions (✓)
●Genome Annotation LiftOver； filter out GC content >=60% (✓)
●Tiling Large Sequences (✓)
Tiling involves breaking up a large region into smaller, overlapping fragments (tiles). This ensures that each tile captures a portion of the regulatory element, which can be tested individually.
●Order lib from company
●Order primers that contain the barcodes
●Synthesize the Library
Additional Background Information:
Massively parallel reporter assays (MPRAs)
High-throughput experimental techniques used to evaluate the regulatory activity of large numbers of DNA sequences, such as enhancers, promoters, and other regulatory elements, simultaneously.
MPRAs allow researchers to test how specific DNA sequences influence gene expression, which is crucial for understanding gene regulation, evolution, and diseases.
Reference:
Gordon MG, Inoue F, Martin B, Schubach M, Agarwal V, Whalen S, et al. lentiMPRA and MPRAflow for high-throughput functional characterization of gene regulatory elements. Nat Protoc. 2020 Aug;15(8):2387–412.
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