The RNA-guided CRISPR-Cas9 nuclease system has revolutionized the genome editing practices. For the most part, the Cas9-mediated genome editing is performed either via nonhomologous end joining (NHEJ) or homology-directed repair (HDR) in mammalian cells, However, designing of specific sgRNAs and minimizing off-target cleavage mediated mutagenesis are the major challenges in CRISPR-Cas based genome editing. To circumvent these issues, we can take advantages of many available tools and approaches for sgRNA construction and delivery.
Get tips on using pSpCas9(BB)-2A-Puro (PX459) V2.0 to perform CRISPR Human - Activation ERαY537S
Get tips on using Q5® Site-Directed Mutagenesis Kit to perform Site Directed Mutagenesis (SDM) Mouse - Point mutation Neuro 2a Epac1
Get tips on using GeneArt™ Site-Directed Mutagenesis System to perform Site Directed Mutagenesis (SDM) Mouse - Point mutation Neuro 2a Epac1
Get tips on using Silencer® Select_FPr1/Silencer® Select_FPr2 siRNA to perform siRNA / miRNA gene silencing Mouse - Neuro 2a Fpr1/Fpr2
Get tips on using NEBNext® Multiplex Small RNA Library Prep Set for Illumina® to perform RNA sequencing Mouse - Neuro 2a
Get tips on using NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina® to perform RNA sequencing Mouse - Neuro 2a
Site-directed mutagenesis (SDM) can be challenging, particularly during detection/confirmation of (SDM) in colonies by sequencing or PCR techniques. This common issue in SDM is heavily relying on designing of mutagenic primer pairs. The best solution is to design the mutagenic primers that have extended 3'-ends/3'-overhang. This would provide the annealing region between the mutagenic primer pair is essentially shorter. and hence ensure a lower annealing temperature for the primer pair along with a higher chance of annealing to the template.
Get tips on using QuikChange II XL Site-Directed Mutagenesis Kit, 10 Rxn to perform Site Directed Mutagenesis (SDM) Mouse - Point mutation Neuro 2a proglucagon
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