Method for knocking out zebrafish slc26a4 gene
A zebrafish and gene technology, applied in the field of gene knockout, can solve the problems of high cost, many steps, time-consuming, reagent and sequencing cost, etc., and achieve the effect of simple production and low cost
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[0043]1) Design CRISPR / Cas9 gene knockout target sites and detection primers
[0044] Query the genomic DNA sequence of the zebrafish slc26a4 gene on the National Center for Biotechnology Information (NCBI), analyze its functional domain on the website SMART (http: / / smart.embl-heidelberg.de / ), and knock out it according to CRISPR / Cas According to the principle, the target site of the slc26a4 gene was designed on the website The ZiFiT Targeter (http: / / zifit.partners.org / ZiFiT / ). The selection of targets must follow this standard: 5'-GG-(N)18-NGG-3'. The GG dinucleotide at the 5' end is part of the T7 promoter, and this restriction is not required when designing the target site, but it must be ensured that the 3' end of the target site is NGG. The selected position of the target must be within the structural domain of the gene to ensure that the insertion or deletion of bases at the target site can affect the entire structural domain of the slc26a4 gene, thereby changing the ex...
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