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polyA Spin mRNA Isolation Kit |
 | | | | polyA Spin mRNA Isolation Kit |
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Prices are in US dollars and valid only for US orders.
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Description:
The polyA Spin™ Kit is a rapid and convenient alternative to traditional column chromatography for the isolation of full-length poly(A)+ (mRNA) from cytoplasmic or "total" RNA isolated from eukaryotic tissue and cells. Poly(A)+ RNA selection is made by affinity chromatography using spin columns. Prepared essentially by the method of Gilham (1), this solid support has found widespread use in the "batch" preparation of low abundance mRNA (2).
Reagents sufficient for the isolation and subsequent precipitation of poly(A)+ RNA from 8 samples of as much as 1.0 mg each of "total" RNA are provided. The isolated RNA can be used for in vitro translation experiments, the preparation of cDNA libraries or Northern analysis.
Advantages:- Prepacked Columns
- 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.
Kit Components:
Elution Buffer
Glycogen Solution
Low Salt Buffer
microcentrifuge spin columns
microcentrifuge tubes containing oligo (dT)25 -cellulose beads
NaAc
sterile microcentrifuge tubes
Wash Buffer
Storage Conditions
Storage Temperature:
4°C
Notes
General notes:- The polyA Spin™ mRNA Isolation Kit complements and serves as an alternative to traditional column chromatography for isolation of mRNA. For total RNA in the 1.0 mg sample range, polyA Spin provides the same resolution as a standard 100 mg column of high quality oligo(dT) cellulose. This is of particular importance for isolation of low abundance messenger RNA. For total RNA samples greater than 1.0 mg, initial "first round" isolation of poly(A)+ material should be done by column (NEB #S1401) chromatography (3). First round isolation of poly(A)+ RNA by either column or spin chromatography typically yields a product contaminated with poly(A)- RNA. polyA Spin™ Kit chromatography is ideal for "second round" purification, yielding material with an A260/A280 of 1.9 or greater with minimal product loss. The protocol provided has been optimized for the isolation for poly(A)+ RNA from 0.5 to 1.0 mg of total RNA isolated from eukaryotic tissue or cells containing between 1-5% mRNA. The amount of poly(A)+ RNA isolated depends on the type of tissue or cells.
Quality Control for Current Lot
Quality control values for a specific lot can be found on the datacard which accompanies each product.
Quality Assurance Statement:
Each lot of the polyA Spin mRNA Isolation Kit is assayed for its ability to isolate mRNA from rat liver total RNA.
References
- Gilham, P.T. (1964) J. Amer. Chem. Soc., 86, 4982.
- Farrell, R.E. (1993) RNA Methodologies, 104-106.
- Jacobson, A. (1987) S. Berger and A. Kimmel (Eds.), Methods Enzymol., 152, pp. 254-257. San Diego: Academic Press.
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