RNA EMSA及问题

  

以下英文内容是我们美国公司网站(www.viagene.com)的原稿内容。因为我们的中文文案常被盗用,就不准备翻译成中文了。如有任何问题,请与我们联系,谢谢理解。

In recent years, evidence has emerged for the existence of many diverse types of RNA, which play roles in a wide range of biological processes in all kingdoms of life. These molecules generally do not, however, act in isolation, and identifying which proteins partner with RNA is a major challenge. Many methods, in vivo and in vitro, have been used to address this question, including combinatorial or high-throughput approaches, such as systematic evolution of ligands, cross-linking and immunoprecipitation and RNA immunoprecipitation combined with deep sequencing. RNA EMSA样图However, most of these methods are not trivial to pursue and often require substantial optimization before results can be achieved. Here, we demonstrate a simple technique that allows one to screen proteins for RNA-binding properties in a gel-shift experiment and can be easily implemented in any laboratory. This assay should be a useful first-pass tool for assessing whether a protein has RNA- or DNA-binding properties, prior to committing resources to more complex procedures.

The principle of RNA EMSA as well as DNA EMSA is simple, considerations for performing a successful RNA EMA including the follows:

1)Needs instruments and reagents for RNA EMSA.

2) The knowledge for handling RNAs and proteins.

3) The skills for running a vertical gel electrophoresis.

4) The facility for imaging ECL signals via x-ray films or imagers for imaging ECL or infrared fluorescence.

5) The RNA probe is expensive, 20 fold of what for making a DNA probes.

6) Most probes for RNA EMSA is a single-strand RNA, which is very fragile for degradation.

7) Working environment should keep as RNase-free.

8) Based on the considerations above, we recommend our IRFluo rEMSA kits for RNA EMSA for its fast results (~2 hours) and simple operation (imaging gels immediately after gel electrophoresis).

9) If using Viagene's RNA EMSA service, the results would be secured as the scientists in Viagene Biotech have rich experience for RNA EMSA (Linking to technique services).

 

【通用RNA EMSA Kits】

产品名 目录号 产品描述 产品包装 价格
CoolShift-BIr SIDET102 非放射性生物素/ECL RNA EMSA成套试剂,(不带探针与膜,查看组份操作说明)。 100次 2860元
CoolShift-IRr SIDET202 非放射性近红外荧光 RNA EMSA成套试剂(不带探针,查看组份操作说明)。 100次 2860元

   

   【RNA EMSA探针定制】

探针类别 目录号 规格 价格 探针用途
生物素RNA探针 BPSRV11
200
IDT RNA合成价+1450元*
用于RNA EMSACompetitive(竞争)与 Supershift (超迁移)EMSA、Pulldown 以及结合蛋白的纯化。
近红荧光RNA探针 BPSRV21
200次
IDT RNA合成价+1750元*
用于常规EMSACompetitive(竞争) EMSASupershift (超迁移)EMSA,可以快速完成测定。
80倍竞争RNA探针 CPSRV11 12次 IDT RNA合成价+20%** 用于Competitive(竞争) EMSA,确定核酸与蛋白结合的特异性。
80倍突变RNA探针 MPSRV21 12次 IDT RNA合成价+20%** 用于Competitive(竞争) EMSA,反证确定核酸与蛋白结合的特异性。
注:*美国IDT网站(www.idtdna.com)查询价格,1450元/1750元的费用包括探针设计、生物素标记或荧光标记、纯化、测定,税费与运费。**20%附加费包括探针设计、订货、税费与运费。

  

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