Protocol detail

Quantifying Genomic Edits via Sanger Sequencing

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Target genomic sites were PCR amplified using the lysate of HEK293T cells and porcine embryos by the Rapid Taq Master Mix (Vazyme), with primers flanking each examined target site. Sanger sequencing was then performed by IGE Biotechnology (Guangzhou, China), and the ratios of C-to-T and A-to-G conversions were calculated by the EditR software^{48 (link)} (https://moriaritylab.shinyapps.io/editr_v10/). The primers used to amplify the target genomic sequences are listed in Supplementary Data 2.

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Chen F., Lian M., Ma B., Gou S., Luo X., Yang K., Shi H., Xie J., Ge W., Ouyang Z., Lai C., Li N., Zhang Q., Jin Q., Liang Y., Chen T., Wang J., Zhao X., Li L., Yu M., Ye Y., Wang K., Wu H, & Lai L. (2022). Multiplexed base editing through Cas12a variant-mediated cytosine and adenine base editors. Communications Biology, 5, 1163.

Publication 2022

Rapid Taq Master Mix

Embryos Genomic Porcine Primers

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Variable analysis

independent variables

Cell type (HEK293T cells and porcine embryos)

dependent variables

Ratios of C-to-T and A-to-G conversions

control variables

Primers flanking each examined target site

controls

No positive or negative controls were explicitly mentioned in the provided information.

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