Journal: iScience

Article Title: Excess of guide RNA reduces knockin efficiency and drastically increases on-target large deletions

doi: 10.1016/j.isci.2023.106399

Figure Lengend Snippet: Key resources table

Article Snippet: ddPCR supermix for probes (no dUTP) , Bio-Rad , 1863024.

Techniques: Recombinant, CRISPR, Staining, Sensitive Assay, SYBR Green Assay, Labeling, Software, Control, Nano Differential Scanning Fluorimetry

Journal: Cold Spring Harbor Molecular Case Studies

Article Title: Somatic IDH1 variant (p.R132C) in an adult male with Maffucci syndrome

doi: 10.1101/mcs.a006127

Figure Lengend Snippet: Sequencing and droplet digital polymerase chain reaction (ddPCR) results. ( A ) Sequence chromatograms showing the somatic IDH1 c.394C > T; p.R132C variant in hemangioma-derived but not blood-derived DNA of an individual with Maffucci syndrome. ( B ) ddPCR readout showing droplets positive (blue) for p.R132C mutant probe in hemangioma-derived but not blood-derived DNA. Droplets for wild-type probe are green and droplets without DNA template are gray. An amplitude of 4000 was set as the positive mutant droplets threshold. Hemangioma-derived DNA: IDH1 p.R132C concentration, 10.24 copies/µL; wild-type concentration, 48.9 copies/µL; mutant allele fraction, 17.3%. Blood-derived DNA: IDH1 p.R132C concentration, 0 copies/µL; wild-type concentration, 138 copies/µL.

Article Snippet: Briefly, probes and primers were mixed with 2× ddPCR Supermix for probe (Bio-Rad) at 217 nM and 435 nM final concentrations for each probe and each of the primers, respectively, and mixed with 10 ng of DNA sample to a final volume of 23 μL.

Techniques: Sequencing, Digital PCR, Variant Assay, Derivative Assay, Mutagenesis, Concentration Assay