Product Class: Kit

polyA Spin™ mRNA Isolation Kit

Product Introduction

This kit contains an affinity matrix used for the isolation of mRNA containing polyadenylic (poly A) regions (1) from purified total RNA. This matrix consists of oligo (dT)25 covalently coupled to a cross-linked cellulose bead. This product is guaranteed for twenty-four months when stored at 4°C in the supplied storage buffer.

  • A rapid and convenient alternative to traditional column chromatography for the isolation of full-length poly(A)+ eukaryotic messenger (mRNA) from samples of Total RNA
  • Shorter purification time: Prepare full-length mRNA in as little as 40 minutes
  • Maintains Resolution: When compared to a 100 mg column of high quality oligo (dT) cellulose in 1.0 mg total RNA sample range
Catalog # Size Concentration
S1560S 8 isolations 0 %

Product Information

Description

The polyA Spin™ Kit is a rapid and convenient alternative to traditional column chromatography for the isolation of full-length poly(A)+ eukaryotic messenger (mRNA) from samples of "total" RNA. Poly(A)+ RNA selection is made by affinity chromatography using spin columns prepackaged with NEB's Oligo (dT)25-Cellulose beads (NEB #S1408S). Prepared essentially by the method of Gilham (1), this solid support has found widespread use in the "batch" preparation of low abundance mRNA (2). Its high binding capacity and rapid hybridization kinetics make it an ideal support for chromatographic poly(A)+ RNA selection. Intact mRNA can be isolated in as little as forty minutes from multiple cell lysates or samples of "total" RNA by spin chromatography in a microcentrifuge.
Reagents sufficient for the isolation and subsequent precipitation of poly(A)+ RNA from eight samples of as much as 1.0 mg of "total" RNA are provided. The isolated RNA can be used for in vitro translation experiments, the preparation of cDNA libraries, Northern analysis, subtractive hybridization or differential display.

Kit Components

The following reagents are supplied with this product:

Store at (°C) Concentration
5 M NaCl 4 500 mM
3M NaAC 300 mM
Wash Buffer I 4 1 X
Wash Buffer (15 ml) 4 1 X
Elution Buffer 1 X
Low Salt Buffer 4 1 X
Glycogen Solution 4 10 mg/ml
microcentrifuge tubes containing oligo (dT)25 -cellulose beads 4
microcentrifuge spin columns
sterile microcentrifuge tubes
Product Categories:
RNA Extraction & Purification
Applications:
RNA Purification and Isolation

Advantages and Features

Features

  • Shorter Purification Time
  • Reduced Isolation Volumes
  • Maintains Resolution: When compared to a 100 mg column of high quality oligo (dT) cellulose in 1.0 mg total RNA sample range.

Properties and Usage

Storage Temperature

4°C

Notes

  1. The polyA Spin mRNA Isolation Kit complements and serves as an alternative to traditional column chromatography isolation of mRNA. Each polyA Spin spin column has the same column performance as a standard 100 mg column of high quality oligo (dT)-cellulose in the 1.0 mg total RNA sample range. Thus the shorter purification times and reduced working volumes associated with spin chromatography are realized while maintaining column chromatography levels of performance. This is of particular importance in the isolation of low abundance messenger RNA (1). For total RNA samples greater than 1.0 mg initial "first round" isolation of poly(A)+ material should be done by column or "batch" (NEB #S1408S) chromatography (2). First round isolation of poly(A)+ RNA by either column or spin chromatography typically yields a product contaminated with poly(A)– RNA. PolyA Spin spin chromatography is ideal for "second round" purification, yielding material with an A260/280 of 1.9 or greater with minimal product loss. The protocol has been optimized for the isolation of poly(A)+ RNA from 0.1–1.0 mg of total RNA isolated from eukaryotic tissue or cells that contain between 1–5% mRNA. The amount of poly(A)+ RNA isolated will vary with the type of tissue or cells used.

References

  1. Gilham, P.T. (1964). J. Amer. Chem. Soc.. 86, 4982.
  2. Farrell, R.E. (1993). RNA Methodologies. 104-106.
  3. Jacobson, A. (1987). S. Berger and A. Kimmel(Ed.), Methods Enzymol.. 152, 254-257. San Diego: Academic Press.

Faqs & Tech Tips

Tech Tips

Do not freeze. To preserve its components’ integrity the kit has to be stored between 2 and 8 degrees.
The denaturing step is critical for adequate mRNA recovery so please, make sure your sample is thoroughly denatured before incubating it with the beads.
Centrifugation speed should never exceed 12,000 x g

Protocols & Manuals

Manuals

The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name these document files: manual[Catalog Number].)

Protocols

  1. Isolation of mRNA Using the PolyA Spin™ mRNA Isolation Kit
  2. Batch Method Protocol (S1560)  for Isolation of mRNA Using the  PolyA Spin™ mRNA Isolation Kit

Other Tools & Resources

Usage Guildelines & Tips

Interactive Tools

Quality & Safety

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.

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality control's 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.