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epicentre MOT60510 mRNA-ONLY™ Prokaryotic mRNA Isol
点击次数：0发布时间：2019/11/14 16:19:02
更新日期：2019/11/14 16:19:02
所 在 地：中国大陆
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详细内容
应用领域
- 分离出基本上不含污染性rRNA的原核mRNA，以及用于RNA扩增和基因表达分析的poly（A）拖尾。
- Isolation of prokaryotic mRNA that is substantially free of contaminating rRNA, ａnd poly(A) tailing for RNA amplification ａnd gene expression analysis.
具有Poly（A）-Tailing *功能的mRNA-ONLY™原核mRNA分离试剂盒提供了一种简单有效的方法：a）分离1小时内基本不含核糖体RNA（rRNA）的原核mRNA；b）分离的原核mRNA的3’-末端poly（A）尾部，用于后续操作或研究。仅mRNA试剂盒包括EPICENTRE的Terminator™5´-磷酸盐依赖性核酸外切酶，*是一种可消化的5´→3´核酸外切酶，可消化具有5´单磷酸的RNA。细菌的rRNA可以作为单转录物转录，然后加工生成具有5＇单磷酸的rRNA，是该酶的底物。因此，具有Poly（A）拖尾的仅mRNA原核mRNA分离试剂盒可用于分离基本不含16S和23S rRNA的原核mRNA。
The mRNA-ONLY™ Prokaryotic mRNA Isolation Kit with Poly(A)-Tailing* provides a simple ａnd effective method for a) isolating prokaryotic mRNA that is substantially free of ribosomal RNA (rRNA) in 1 hour; ａnd b) 3´-end poly(A)-tailing of the isolated prokaryotic mRNA for subsequent manipulation or study. The mRNA-ONLY Kit includes EPICENTRE＇s Terminator™ 5´-Phosphate-Dependent Exonuclease,* a processive 5´→3´ exonuclease that digests RNA having a 5´ monophosphate. Bacterial rRNAs, which are transcribed as a single transcript ａnd then processed to yield rRNAs with 5＇ monophosphates are substrates for the enzyme. Thus, the mRNA-ONLY Prokaryotic mRNA Isolation Kit with Poly(A) Tailing can be used to isolate prokaryotic mRNA substantially free of 16S ａnd 23S rRNA.
仅mRNA试剂盒还包括EPICENTRE的Poly（A）聚合酶，该酶使用ATP作为底物，将模板磷酸单磷酸腺苷独立地添加到RNA分子的3′-羟基末端。从总RNA混合物中去除rRNA大大提高了poly（A）拖尾反应的效率（图1）。然后将所得的聚腺苷酸化的mRNA用于下游应用，例如cDNA合成和基因表达分析。
The mRNA-ONLY Kit also includes EPICENTRE＇s Poly(A) Polymerase which uses ATP as a substrate for template-independent addition of adenosine monophosphates to the 3´-hydroxyl termini of RNA molecules. Removal of rRNA from the total RNA mixture greatly increases the efficiency of the poly(A)- tailing reaction (Fig. 1). The resulting polyadenylated mRNA can then be used for downstream applications like cDNA synthesis ａnd gene expression analysis.
优势说明
- 包括聚（A）尾剂；无需单独购买。
- Poly(A)-tailing reagents included; no separate purchase required.
- The kit can be used to simultaneously prepare mRNA from mixed populations of prokaryotes or from mixed populations of both prokaryotic ａnd eukaryotic cells, such as occur in biological samples involving symbiotic, parasitic, or pathogenic interactions (e.g., nitrogen fixation, prokaryotic infections, biological films, etc.).
- 该试剂盒可用于同时从原核生物的混合种群或原核和真核细胞的混合种群制备mRNA，例如存在于涉及共生，寄生或致病相互作用（例如固氮，原核感染，生物膜）的生物样品中等）。
Figure 1. Terminator™ Exonuclease ａnd Poly(A) Polymerase treatment of total bacterial RNA spiked with a tracer transcript. E. coli total RNA (lane 1) ａnd a 718-nt transcript were combined ａnd then treated with Terminator Exonuclease to remove rRNA (lane 3). The rRNA-free sample was then treated with Poly(A) Polymerase for poly(A)-tailing of the mRNA (lane 4). An untreated mix of total RNA ａnd the tracer (lane 5) was also treated only with Poly(A) Polymerase (lane 6). The tracer is clearly tailed more efficiently in the absence of rRNA. Lane 2, MW marker.
