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mRNA-ONLY™ Prokaryotic mRNA Isolation Kit with Poly(A)-Tailing

货号

MOT60510

规格

10 Rxns

存储温度

-20℃

产品详情

Applications

Isolation of prokaryotic mRNA that is substantially free of contaminating rRNA, and poly(A) tailing for RNA amplification and gene expression analysis.

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.

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.

Benefits

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.).

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.

*Covered by issued and/or pending patents.

Catalog No.	Size

mRNA-ONLY™ Prokaryotic mRNA Isolation Kit with Poly(A)-Tailing
MOT60510	10 Reactions
Contents: Terminator™ 5´-Phosphate-Dependent Exonuclease, mRNA-ONLY™ Prokaryotic 10X Reaction Buffers A and B, RiboGuard™ RNase Inhibitor, mRNA-ONLY™ Stop Solution, 5M LiCl Solution, Poly(A) Polymerase, 10X Poly(A) Polymerase Reaction Buffer, ATP, RNase-Free Water.

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