CRISPR-Chip for Detecting BFP and DMD

For bfp detection via CRISPR–Chip, target HEK-BFP and non-target HEK genomic materials lacking the bfp sequence were utilized. Briefly, HEK-BFP DNA was extracted from bfp-transduced HEK293T cells (PureLink Genomic DNA Mini Kit; Thermo Fisher Scientific) and purified (QIAquick PCR Purification Kit; Qiagen) according to the manufacturer’s instructions and published protocol^{36 (link)}. For analysis of DMD-associated mutations via CRISPR–Chip, human genomic samples from both healthy male and DMD male patients were purchased with certificate of analysis from the Coriell Institute for Medical Research. DMD samples presented in this study carried previously identified large-scale fragment deletions: A (NA07691)^{44 (link)}, B (NA03780)^{44 (link)}, C (NA03782)^{44 (link)}, D (NA04100)^{44 (link)}, E (NA05126)^{44 (link)} and F (NA04364)^{44 (link)}. Healthy samples—H1 (NA22264), H2 (NA22807) and H3 (NA03798)—were also validated for the presence of target exons (see Supplementary Fig. 14). The concentration of genomic material was routinely measured before incubation with CRISPR–Chip using Nanodrop (Infinite M200 NanoQuant; Tecan).

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Hajian R., Balderston S., Tran T., deBoer T., Etienne J., Sandhu M., Wauford N.A., Chung J.Y., Nokes J., Athaiya M., Paredes J., Peytavi R., Goldsmith B., Murthy N., Conboy I.M, & Aran K. (2019). Detection of unamplified target genes via CRISPR–Cas9 immobilized on a graphene field-effect transistor. Nature biomedical engineering, 3(6), 427-437.

Publication 2019

PureLink Genomic DNA Mini Kit QIAquick PCR Purification Kit Infinite M200 NanoQuant

Chip Crispr Deletions Exons Genomic Genomic human M200 Male Male human Mutations Patients

Corresponding Organization :

Other organizations : Keck Graduate Institute, Claremont Colleges, University of California, Berkeley, Universidad de Navarra

Top 5 similar protocols

Variable analysis

independent variables

Target HEK-BFP genomic materials
Non-target HEK genomic materials lacking the bfp sequence
Human genomic samples from both healthy male and DMD male patients

dependent variables

Bfp detection via CRISPR–Chip
Analysis of DMD-associated mutations via CRISPR–Chip

control variables

Concentration of genomic material measured before incubation with CRISPR–Chip

controls

Positive control: HEK-BFP DNA extracted from bfp-transduced HEK293T cells
Negative control: Non-target HEK genomic materials lacking the bfp sequence
Healthy genomic samples (H1, H2, H3) validated for the presence of target exons

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