Multiplexed Capture and Sequencing of Select Genes
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Corresponding Organization :
Other organizations : The Ohio State University, Nationwide Children's Hospital, Case Western Reserve University, Massachusetts General Hospital, Centro Clinico Nemo, University of Milan
Variable analysis
- Captured and sequenced genes of interest using the Multiplexed Direct Genomic Selection (MDiGS) assay
- Used specific BAC clones for capture: RP11-652K3 (CFTR), RP11-1056O6 (CFTR), RP11-268A15 (PLS3), and a 35.5 kb portion of RP1-215P15 containing the entire SMN2 gene
- Sequences of the captured genes of interest (CFTR, PLS3, SMN2)
- Used the same method as described in Alvarado et al. (2014) for the MDiGS assay
- Pooled 48 indexed DNA samples
- Used four cosmids (108F4, 121C9, 22A5, 30C9) flanking the SMN2 gene to block non-specific capture, and confirmed they do not contain SMN2 by PCR
- Positive control: Not explicitly mentioned.
- Negative control: The four cosmids flanking the SMN2 gene were used as a negative control to ensure specificity of capture.
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