CFTR Long Read Sequencing Protocol using Oxford Nanopore Technology v1

Cystic fibrosis (CF) is a genetic autosomal recessive disorder, commonly associated with European populations, with insufficient awareness in African populations. Whole-gene CFTR sequencing is effective for identifying diverse CFTR variants, facilitating accurate CF carrier screening in African populations. The proposed method uses long-read Oxford Nanopore Technologies to sequence the entire CFTR gene. This involves amplifying ~25Kb fragments of the CFTR gene using specially designed long-range PCR primer pairs, followed by barcoded library preparation and sequencing with ONT's R10.4 flow cells and the Mk1C device. This approach is especially advantageous for genetic laboratories with limited resources, due to its cost-effectiveness and relative ease of implementation.