Isolating high-quality RNA molecules is crucial to many downstream experiments, such as cloning,reverse transcription for cDNA library creation, sequencing and structural analysis. Studies that require the isolation of eukaryotic mRNA often take advantage of the polyadenine tail typically present at the 3’ end. One approach involves the use affinity matrices; poly-T oligonucleotides that hybridize to the complementary poly-A tails (1). Enrichment of miRNA and siRNAs can also be achieved using high-affinity binding proteins and the use of chitin magnetic beads (2, 3).
(1) Aviv, H. and Leder, P. (1972) Proc. Natl. Acad. Sci. USA, 69, 1408. PMID: 4504350
(2) Silhavy, D. et al. (2002) EMBO J., 21, 3070-3080. PMID: 12065420
(3) Jin, J. et al. (2010) Biotechneques. 48, XVII-XXII. PMID: 20569217
Protocols for RNA Isolation & Detection
- Quantitation of Isolated Poly(A)+ RNA: From the Magnetic mRNA Isolation Kit
- Batch Method Protocol (S1560)
- Isolation of mRNA from Mammalian Cells: Using The Magnetic mRNA Isolation Kit
- Isolation of mRNA from Tissue: Using the Magnetic mRNA Isolation Kit.
- Isolation of mRNA (S1560)
- Ligation protocol using SplintR® Ligase (M0375)
- Protocol for isolation of siRNA from p19 siRNA Binding Protein (M0310) using chitin magnetic beads
- Streptavidin Magnetic Beads
- cDNA synthesis in oligo (dT)25 magnetic beads (S1419)
- Phage Display: Solution-phase Panning with Affinity Bead Capture
- Avoiding Ribonuclease Contamination
Other Tools & Resources
Reagents for RNA Sample Preparation
RNA Polymerase Selection Chart
RNA Ligase Selection Chart
cDNA Synthesis Selection Chart
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