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- 详细信息
- 文献和实验
- 技术资料
- 英文名:
sgRNA In Vitro one-step Transcription Kit
- 规格:
10次
目录号:E369-01A
产品描述:
CRISPR/Cas9是一种来源于细菌获得性免疫的由RNA指导Cas9蛋白对靶向基因进行编辑的技术。CRISPR/Cas9的切割是通过向导RNA(gRNA)与打靶基因组位置之间约20bp碱基的配对,且打靶序列的3′端需要有PAM结构(NGG),再引导Cas9作用实现的。CRISPR/Cas9与靶位点识别的特异性主要依赖于gRNA与靠近PAM处10~12bp碱基的配对,而其余远离PAM处8~10bp碱基的错配对靶位点的识别影响不明显,这导致了CRISPR/Cas9存在一定的脱靶性,可能发生非特异性切割,引起基因组非靶向位点的突变,这样会造成研究结果的不确定性以及研究工作量的大量增加。
Novoprotein CRISPR/Cas9基因编辑系统能有效解决这一问题。sgRNA体外转录试剂盒(Cat.No: E369 专利号ZL 2018 1 0634615.8)30分钟一步法合成sgRNA,再通过sgRNA体外筛选试剂盒(Cat.No:E370)筛选出效率高的sgRNA,用于Crispr基因编辑实验。
产品特点:
30分钟,一步法高效转录sgRNA,操作简便快捷。
sgRNA产量可达10-30ug。
产品用途:
为CRISPR/Cas9基因编辑系统高效转录sgRNA。
保存温度:
-20℃保存,或-80℃长期保存。
应用实例:
图例)体外转录97bp sgRNA片段,并作体外切割实验。

专利证书:
![2018-09-25.[P2018-1147].åæä¸å©è¯ä¹¦.jpg](https://www.novoprotein.com.cn/Public/Uploads/ueditor/upload/image/20181018/1539828962135407.jpg)
参考文献:
Virus-like nanoparticle as a co-delivery system to enhance efficacy of CRISPR/Cas9-based cancer immunotherapy[J]. Qi Liu. et al. BIOMATERIALS. 2020.
An interferon-independent lncRNA promotes viral replication by modulating cellular metabolism[J]. Pin Wanget al. Science. 2017.
An in vitro site-specific cleavage assay of CRISPR-Cas9 using a personal glucose meter[J]. Shaohua Gong. et al. Chemical Communications. 2020.
CRISPR/Cas9-mediated editing of CsWRKY22 reduces susceptibility to Xanthomonas citri subsp. citri in Wanjincheng orange (Citrus sinensis (L.) Osbeck)[J]. Lijuan Wang et al. Plant Biotechnology Reports. 2019.
For research use only.
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文献和实验The following protocol is for MEGAscript II Kit (Ambion). 1. PCR amplify the DNA template. The 5'-end of the template should contain the minimum promoter sequenences of T7 or Sp6 or T3. A 5-primer with the minimum promoter sequences may be used
An RNA Polymerase II In Vitro Transcription System
A great deal of attention in recent years has been focused on the mechanisms governing transcription. Repression and/or activation of specific genes, or sets of genes, represents a key regulatory step in such diverse processes as cell growth
In vitro transcription with yeast nuclear extract
In vitro transcription with yeast nuclear extract Steve Hahn Last Modified Fri, Apr 25, 2003 Wear gloves throughout, use RNAse free solutions (either autoclaved or sterile filtered) and clean bench and pipetmen with 95% ethanol before use
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