The New England Biolabs Magnetic mRNA Isolation Kit is designed to isolate intact poly(A)+ RNA from cells and tissue without requiring phenol or other organic solvents. The technology is based on the coupling of Oligo d(T)25 to 1 μm paramagnetic beads which is then used as the solid support for the direct binding of poly(A)+ RNA. Thus, the procedure permits the manual processing of multiple samples and can be adapted for automated high-throughput applications. Additionally, magnetic separation technology permits elution of intact mRNA in small volumes eliminating the need for precipitating the poly(A)+ transcripts in the eluent. Intact poly(A)+ RNA which is fully representative of the mRNA population of the original sample can be obtained in less than one hour. Oligo d(T)25 Magnetic Beads can be reused up to three times and the researcher has the option of eluting the isolated mRNA or using the bound dT DNA as a primer in a first-strand cDNA reaction. Figure 1:
Consistency and wide isolation range are demonstrated by poly(A)+ RNA isolation from duplicate samples of decreasing numbers of HEPG2 cells (5 x 105 to 1 x 103) by direct lysis/binding in microtiter plate followed by mRNA isolation with the magnetic method. 1/10th of isolated mRNA is converted to oligo (dT) primed cDNA using ProtoScript First Strand cDNA Synthesis Kit (NEB #E6500) and qPCR done with validated primers for the peptidylpropyl isomerase, a low-abundance housekeeping gene.
Figure 2:
Facilitate the isolation of poly(A)+ RNA using the Magnetic mRNA Isolation kit
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