RNA 2 or BRNA 2 were annealed with RNA 5 to generate non biotinylated or biotinylated dsRNA; RNA 2 or BRNA 2 were annealed with DNA 9 to generate nonbiotinylated or biotinylated RNA:DNA hybrids; DNA 9 or BDNA 9 were annealed with DNA 6 to obtain nonbiotinylated or biotinylated dsDNA. Annealing was performed in a MasterCycler nexus gradient (Eppendorf) in annealing buffer [60 mM NaCl, 5 mM tris-HCl (pH 7.5), and 0.2 mM EDTA] using the following program: 4 min at 95°, 85°, 82°, 80°, 78°, 75°, and 72°C, followed by 10 min at 70°C and slow cooling down to 10°C over a period of 2 hours. The quality of annealed material was controlled by running 500 ng of annealed material on a 10% nondenaturing acrylamide gel using ssRNA and ssDNA as controls.

RNA 2: UUU CAA UUC CUU UUA GGA UUA AUC UUG AAG AUA GAG UUA A

RNA 5: UUA ACU CUA UCU UCA AGA UUA AUC CUA AAA GGA AUU GAA A

DNA 9: TTA ACT CTA TCT TCA AGA TTA ATC CTA AAA GGA ATT GAA A

DNA 6: TTT CAA TTC CTT TTA GGA TTA ATC TTG AAG ATA GAG TTA A

BRNA 2: BIO- UUU CAA UUC CUU UUA GGA UUA AUC UUG AAG AUA GAG UUA A

BDNA 9: BIO- TTA ACT CTA TCT TCA AGA TTA ATC CTA AAA GGA ATT GAA A

注意:以上内容是从某篇研究文章中自动提取的,可能无法正确显示。



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