Isolation of Functional RNA from Periderm Tissue of Potato Tubers and Sweet Potato Storage Roots
Tóm tắt
A reliable and efficient protocol is given for the isolation of mRNA from the periderm of potato tubers and sweet potato storage roots. The method relies on a urea-based lysis buffer and lithium chloride to concentrate total RNA away from most of the cytoplasmic components and to prevent oxidation of phenolic complexes. To enhance the physical separation of the RNA from other macromolecular components, the RNA fraction was incubated in the presence of the cationic surfactant Catrimox-14. Poly(A)+ mRNA was separated from total RNA and other contaminants by using Promega's MagneSphere technology. The mRNA was suitable for cDNA library construction and RNA fingerprinting.
Tài liệu tham khảo
Ahmad F and Bigelow CC (1979) The denaturation of ribonuclease A by combinations of urea and salt denaturants. J Mol Biol 13: 607-17.
Bugos RC, Chiang VL, Zhang XH, Campbell ER, Podila GK and Campbell WH (1995) RNA isolation from plant tissues recalcitrant to extraction in guanidine. Biotech 19: 734-737.
Chomczynski PA and Sacchi N (1991) Single-step isolation from cultured cells or tissue. Unit 4.2 (Supplemented 14). In: Current Protocols in Molecular Biology. Ausubel FM, Brent R, Kingston RE, Moore DD, Smith JA, Seidman JG and Struhl K (eds), John Wiley & Sons, New York.
Hughes DW and Galau GA (1988) Preparation of RNA from cotton leaves and pollen. Plant Mol Biol Rep 6: 253-275.
Johnson ML, Redmer DA and Reynolds LP (1995) Quantitation of lane-to-lane loading of Poly(A)RNA using a biotinylated oligo (dT) probe and chemiluminescent detection. Biotech 19: 712-714.
Levi A, Galau G and Wetzstein HY (1992) A rapid procedure for the isolation of RNA from high-phenolic-containing tissues of pecan. Hortsci 27: 1316-1318.
Lewinsohn E, Steele CL and Croteau R (1994) Simple isolation of functional RNA from woody stems of gymnosperms. Plant Mol Biol Rep 12: 20-25.
Logemann J, Schell J and Willmitzer L (1987) Improved method for isolation of RNA from plant tissue. Anal Biochem 163: 16-20.
Macfarlane DE and Dahle CE (1993) Isolating RNA from whole blood-the dawn of RNA-based diagnosis. Nature 362: 186-188.
Smale G and Sasse J (1992) RNA isolation from cartilage using density gradient centrifugation in cesium triflouroacetate: An RNA preparation technique effective in the presence of high proteoglycan content. Anal Biochem 203: 352-356.
