RNase 4 Digestion and 3´ End Repair Mix
Product informationCode | Name | Size | Quantity | Price | |
---|---|---|---|---|---|
M1288S |
RNase 4 Digestion and 3′ End Repair Mix |
50 rxns | - | Unavailable in your region | |
M1288L |
RNase 4 Digestion and 3′ End Repair Mix |
250 rxns | - | Unavailable in your region |
RNase 4 Digestion and 3´ End Repair Mix
Catalog #M1288
Product Introduction

RNase 4 Digestion and 3′ End Repair Mix is a coformulation of RNase 4 (cut sites: U/A and U/G) and T4 Polynucleotide Kinase that simplifies RNase 4 digestion products into a pool of U-ending RNA oligonucleotides with a homogeneous 3′-hydroxy termini.
- Generate a larger population of unique and easy-to-map oligonucleotides for improved RNA characterization by LC-MS/MS.
- Endoribonuclease activity tolerates common RNA chemical modifications.
- For mRNA 5´ cap analysis, use stand-alone RNase 4 with our protocol for DNA Probe-Directed Analysis of mRNA 5´Cap Structures
- Product Information
- FAQs & Troubleshooting
- Citations & Technical Literature
- Quality, Safety & Legal
- Other Products You May Be Interested In
Product Information
Description
RNase 4 is a single-stranded RNA endonuclease that cleaves 3´ of uridine in uridine-purine sequences (cut sites: U/A and U/G). Digestion of RNA with RNase 4 produces U-ending RNA oligonucleotides that contain cyclic 2´, 3´-phosphate and/or linear 3´-phosphate termini. RNase 4 Digestion and 3′ End Repair Mix is a coformulation of RNase 4 and the 3´ end repair activity of T4 Polynucleotide Kinase to produce a pool of U-ending RNA oligonucleotides with a 3´-hydroxy terminus. The additional 3´ end repair of RNase 4 digestion products simplifies and improves RNA sequencing coverage analysis and modification mapping by liquid chromatography-mass spectrometry (LC-MS/MS). RNase 4 endoribonuclease activity tolerates uridine base modifications such as pseudo-, N1-methyl-pseudo-, dihydro-, and 5-methoxy-uridine species (Ψ, m1Ψ, D, and mo5U).
Sequence: To verify the mRNA sequence and its modification status, samples are digested with RNase 4 to produce a series of defined oligonucleotides in sizes that are amenable to LC-MS/MS analysis. A coverage map can be generated by aligning the observed oligonucleotides to the reference sequence. RNase 4 Digestion and 3´ End Repair Mix simplifies mapping by producing a single pool of RNA digestion products with homogeneous 3´ hydroxy termini. One or more additional RNases (e.g., RNase T1) can be used in parallel digestions to increase the total coverage of mRNA.



- This product is related to the following categories:
- RNases,
- RNA Modification,
Reagents Supplied
Reagents Supplied
The following reagents are supplied with this product:
NEB # | Component Name | Component # | Stored at (°C) | Amount | Concentration | |
---|---|---|---|---|---|---|
Properties & Usage
Reaction Conditions
1X NEBuffer™ r1.1
Incubate at 37°C
1X NEBuffer™ r1.1
10 mM Bis-Tris-Propane-HCl
10 mM MgCl2
100 µg/ml Recombinant Albumin
(pH 7 @ 25°C)
Related Products
Companion Products
References
- Wolf et al. (2022). Human RNase 4 improves mRNA sequence characterization by LC-MS/MS. NAR. PubMedID: 35871301
FAQs & Troubleshooting
FAQs
- How much RNase 4 Digestion and 3′ End Repair Mix (NEB #M1288) should I use to digest RNA for LC-MS/MS nucleotide sequence mapping?
- Does RNase 4 (NEB #M1284) cut RNA at sites other than U/A and U/G?
- Can I use RNase 4 Digestion and 3′ End Repair Mix (NEB #M1288) for DNA-probe directed 5′ cap mRNA analysis?
- Can RNase 4 (NEB #M1284) be inactivated?
- Is RNase 4 Digestion and 3′ End Repair Mix (NEB #M1288) sensitive to RNA modifications?
- Does RNA secondary structure impact RNase 4 (NEB #M1288) activity?
- Does RNase 4 Digestion and 3′ End Repair Mix (NEB #M1288) require magnesium?
- Can you tell me more about the switch from BSA to Recombinant Albumin (rAlbumin) in NEBuffers?
Citations & Technical Literature
Citations
Quality, Safety & Legal
Quality Assurance Statement
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]Certificate Of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]Safety DataSheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.RNase 4 Digestion and 3´ End Repair Mix
NEBuffer™ r1.1
Legal and Disclaimers
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
No supporting documents available
This product has no supporting documents available for download. If you feel like supporting documents should be available for this product, please contact us.