versadoc tm mp 4000 system  (Bio-Rad)

Journal: International Journal of Molecular Sciences

Article Title: An Influence of Modification with Phosphoryl Guanidine Combined with a 2′-O-Methyl or 2′-Fluoro Group on the Small-Interfering-RNA Effect

doi: 10.3390/ijms22189784

Figure Lengend Snippet: The products of siRNA duplex degradation after 7.5 µg/mL RNAse A treatment, according to denaturing PAGE. ( A ). The black arrow indicates cleavage sites C 4 identified in an FITC-labeled antisense strand; red arrow: U 12 in an FITC-labeled antisense strand; green arrow: the C 16 or U 17 site in an FITC-labeled antisense strand; blue arrow: U 18 in the unlabeled sense strand of the duplex. ( B – F ): Gel images of 20% PAAGs with corresponding FITC-labeled antisense chains; +RNase A : duplexes treated with Rnase A; -RNAse A : control (undigested) duplexes. Each sample contained 32 P-labeled and FITC-labeled antisense chains of the same structure (1.5 and 50 pmol, respectively) and 55 pmol of an unlabeled sense chain. Antisense strands are designated at the bottom of each image ( B: As, C: As-Ome, D: As-Ome*, E: As-F, and F: As-F* ). Lane 1 contains a native sense ( S ) chain in all images. Other lanes contain a modified sense chain: Lane 2 : S-F, Lane 3: S-F*, Lane 4 : S-Ome , and Lane 5 : S-Ome *. The products of degradation of FITC-labeled siRNAs duplexes were visualized in the gel via scanning and recording of an image using a VersaDoc TM MP 4000 Molecular Imager ® System (Bio-Rad) after laser excitation at 488 nm. Lanes LB and T1 present the stochastic hydrolysis in 50 mM NaHCO 3 –Na 2 CO 3 buffer (pH 9.5) and partial digestion of FITC-labeled As with Rnase T1, respectively. The asterisk (*) means the PG-modified chain.

Article Snippet: After electrophoretic separation, the products of degradation of the FITC-labeled siRNA duplexes were visualized in the gel by scanning and recording an image by means of the VersaDoc TM MP 4000 Molecular Imager ® System (Bio-Rad) after laser excitation at 488 nm.

Techniques: Labeling, Modification

Journal: International Journal of Molecular Sciences

Article Title: An Influence of Modification with Phosphoryl Guanidine Combined with a 2′-O-Methyl or 2′-Fluoro Group on the Small-Interfering-RNA Effect

doi: 10.3390/ijms22189784

Figure Lengend Snippet: Cleavage site determination in FITC-labeled modified siRNAs by denaturing PAGE. Electropherograms for products of degradation of five siRNAs after 7.5 µg/mL RNAse A treatment. Each image has a D lane representing an RNAse A-treated duplex that contains S-OMe* and various derivatives of the antisense chain ( As-F* , As-F, As-OMe* , As-OMe , or As ). The products of degradation of FITC-labeled siRNA duplexes were visualized in the gel via scanning and recording of an image by means of the VersaDoc TM MP 4000 Molecular Imager ® System (Bio-Rad) after laser excitation at 488 nm. Lanes LB and T1 are the stochastic hydrolysis in 50 mM NaHCO 3 –Na 2 CO 3 buffer (pH 9.5) and partial digestion of the FITC-labeled As with RNase T1, respectively. The asterisk (*) means the PG-modified chain.

Article Snippet: After electrophoretic separation, the products of degradation of the FITC-labeled siRNA duplexes were visualized in the gel by scanning and recording an image by means of the VersaDoc TM MP 4000 Molecular Imager ® System (Bio-Rad) after laser excitation at 488 nm.

Techniques: Labeling, Modification