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包装
kit of 6 vials (reagents for 25 Reactions)
运输
dry ice
储存温度
−20°C
一般描述
Following a gene editing experiment, genomic DNA surrounding the target locus is amplified by PCR, and the PCR amplicons are denatured and reannealed through heating and slow cooling. If NHEJ events have occurred, then, after reannealing, several products are possible. Homoduplexes can form where a WT strand is reannealead to a WT strand or an indel-carrying strand is reannealed to an indel-carrying strand. Heteroduplexes form when a WT strand is reannealed to an indel-carrying strand causing a mismatch. Heteroduplex products with mismatches are cleaved by the T7 endonuclease. Separating the DNA products after treatment with T7 endonuclease by gel electrophoresis will result in a banding pattern indicative of the amount of heteroduplexes in the sample. The amount of cleaved heteroduplexes is directly related to the amount of indel activity.
应用
特点和优势
- Technically simple method based on well-known techniques
- Easily interpretable results
- Fast analysis turnaround
- Cost-effective
组分
- one vial of T7 Endonuclease I
- one vial of Control Template and Primer Mix
- one vial of Buffer solution
- one vial of DNA Ladder - 1KB
- one vial of Gel Loading Dye (6X)
- one vial of Proteinase K
原理
Efficiency in gene editing can vary in large part due to the target sequences. Chromatin structure and some sequence elements, for example high GC-content, can inhibit editing at some genomic sequences, affecting sgRNA activity. Additionally, favorable bases in the sgRNA sequence such as a guanine proximal to the PAM can promote sgRNA activity, but these preferred bases may not be available at the target site. It is important to evaluate the gene editing ability of several sgRNAs by quantifying the frequency of modifications using a method like T7 endonuclease mismatch detection.
商品
Validate CRISPR gene editing experiments easily with Sigma-Aldrich® T7E1 mismatch detection kit, ensuring successful editing.
实验方案
Guaranteed PURedit™ CRISPR synthetic gRNAs and Cas9 protein offer industry-leading on-site cutting and specificity
可靠的PURedit™ CRISPR合成gRNA和Cas9蛋白可提供行业领先的靶位点切割和特异性
Combine guaranteed sgRNAs with our comprehensive range of CRISPR products and tools, including Cas9 and delivery reagents, for efficient genome modification with higher specificity.
相关内容
超丰富的优质 Cas9 蛋白组合,确保您始终可找到满足自身特定实验需求的完美 CRISPR 试剂。严苛纯化、NLS 信号、增强特异性和活性,确保您高效、简便地开展基因编辑实验。
Quality Cas9 proteins ensure efficient gene editing with increased specificity and activity, catering to diverse CRISPR reagent needs.
Explore technology and reagent portfolios for plant breeding workflows, accelerating the development of new crop varieties.
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