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[…] od in a qPCR machine or end-point fluorescence reading in a plate reader after regular PCR. With this versatility, ASQ may prove to be the method of choice for rapid, accurate, and cost-effective genotyping., All the reagents and instruments used in our experiments are listed in (Supplementary Data are available online at, Nucleotide sequences were reviewed with Sequencher (ver. 5.3; Gene Codes Corporation, Ann Arbor, MI). The determination of annealing temperatures of the primers was based on SnapGene (ver. 2.8.2; GSL Biotech LLC, Chicago, IL). Potential secondary structures, including hairpin, self-dimers, and heterodimers of oligo nucleotides, were reviewed with OligoAnalyzer (ver. 3.1; IDT Inc., Coralville, IA). Nonspecific amplification of fluorescent probes and primers was reviewed with Primer-BLAST (National Center for Biotechnology Information, Bethesda, MD). Fluorescence and Ct (threshold cycle) values were quantified and converted to raw data with CFX Manager (Bio-Rad Laboratories, Inc., Hercules, CA)., All zebrafish strains were maintained and handled following standard practices and guidelines from the Institutional Animal Care and Use Committee (IACUC) in Mayo Clinic. myo7aa strain was provided by the Nicolson laboratory. All other strains were made in our laboratory using TALENs targeting the gene of interest (nr3c1, nod2, mc2r, and il-6). The GoldyTALEN scaffolding was used to construct TALENs., An established hot sodium hydroxide (NaOH) DNA extraction method was used. The hot NaOH DNA extraction allowed a rapid preparation of DNA in 1 hr without additional purification. Although it was a crude preparation, the DNA preparation co […]

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