Poly(A) Polymerase, E. coli
Cat. No.
E099
Unit
25 U (25 µl)
Price
$129.00
| Cat. No. | E099 | ||||||||
| Name | Poly(A) Polymerase, E. coli | ||||||||
| Unit | 25 U (25 µl) | ||||||||
| Category | Molecular Biology Enzymes and Kits | ||||||||
| Description |
Poly(A) Polymerase, E. coli (E-PAP) catalyzes the template-independent addition of adenosine residues to the 3’ ends of RNA, efficiently creating a poly(A) tail. Unlike yeast-derived enzymes, E-PAP works in a sequence-independent manner, meaning it does not require specific recognition sequences to function. This allows for rapid polyadenylation at virtually any unprotected 3’ RNA terminus, regardless of secondary structure. E-PAP is ideal for applications that require poly(A) tail addition to a wide range of RNA molecules, including RNA sequencing and cDNA synthesis. Its versatility and efficiency make it a valuable tool for researchers working with RNA in various experimental setups.
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| Application |
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| Concentration | 1U/µl | ||||||||
| Components | Enzyme is supplied with a 10X Reaction Buffer | ||||||||
| Storage Buffer | 20mM Tris-HCl, 100mM NaCl, 0.1mM DTT, 0.1mM EDTA, 50% Glycerol, 0.1% Triton X-100, pH 7.5 @25°C | ||||||||
| Storage Condition | Store all components at -20°C. | ||||||||
| Note |
One unit is defined as the amount of enzyme that will incorporate 1 nmol of AMP into RNA in a 20 µl volume in 10 minutes at 37°C. |
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| Caution | This product is distributed for laboratory research only. Not for diagnostic use. | ||||||||
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. E099 |
Datasheet
| How long is the typical poly(A) tail generated using this enzyme? | |
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The Poly(A) Polymerase, E. coli can generate a poly(A) tail of over 100 bases when 1 unit of enzyme is incubated with 1–10 µg of RNA in a 20 µl reaction at 37°C for 30 minutes.
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| Does this enzyme require a specific sequence for polyadenylation? | |
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No, unlike Poly(A) polymerase, Yeast, this E. coli enzyme does not require specific recognition sequences and can rapidly add poly(A) tails to nearly all unprotected 3’ RNA termini.
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| How can I stop the reaction after incubation? | |
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The reaction should be stopped by immediately proceeding to an RNA clean-up step. abm recommends using their RNA Purification Magnetic Beads (Cat. No. G971) for best results.
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| What factors influence the length of the poly(A) tail? | |
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Several factors can affect the tail length, including RNA 3’ hydroxyl end concentration, incubation time, ATP concentration, and the amount of enzyme used.
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| Can this enzyme be used with structured RNA molecules? | |
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Yes, this enzyme is effective regardless of RNA secondary structure, as it does not require specific sequences for activity.
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| What are the key differences between Yeast and E. coli Poly(A) Polymerase? | |||||||||||||||||||
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| What is the source of this enzyme? | |
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The enzyme is produced recombinantly in E. coli, which has been engineered to express the enzyme gene. While the original gene may come from another organism, all production and purification occur using E. coli under controlled conditions.
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