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Here, we report a multifunctional mutagenesis system in zebrafish using the maize Ds transposon.
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Sleeping Beauty (SB) is a genetically engineered insertional mutagenesis system.
However, because of potential polar effects of the TargeTron insertion, we developed a CRISPR-Cas9 mutagenesis system for C. difficile.
Site-directed mutagenesis of selected residues for SlTRMs and OVATE/SlOFP20 was carried out with the QuikChange II XL Site-Directed Mutagenesis system (Agilent Technologies, Santa Clara, CA) according to the manufacturer's specifications.
All the mutagenesis of B56γ1 and BubR1 mentioned in this paper were made by Fast mutagenesis system kit (Transgen) and purified by the same way as wild type.
The G608G mutation at exon 11 was introduced using the GeneTailor Site-Directed Mutagenesis System (Invitrogen).
Mutagenesis of alsSD and nuc was done as previously described [52] using the pKOR1 mutagenesis system [45].
Site-directed mutagenesis was performed using the GeneTailor Site-Directed Mutagenesis System (Invitrogen Corporation) according to the manufacturer's recommendations.
We generated hos1/hos1, hos2/hos2, and rpd31/rpd31 mutants using the Tn7::UAU1 insertional mutagenesis system [24].
Point mutations/deletions were generated in the constructs using the Quickchange site-directed mutagenesis system (Strategene).
Based on this, we independently generated an alsSD mutant UAMS-14899) using the pKOR1 mutagenesis system [45].
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