polyA Spin™ mRNA Isolation Kit
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
Avoiding RNase Contamination
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.
The following reagents are supplied with this product:
Store at (°C) Concentration 5 M NaCl 4 500 mM 3M NaAC 4 300 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
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