1 uM “Sub_Ebo” RNA was reacted with XNAzyme “FR6_1” under high magnesium selection conditions (20 h, 17 °C) and the 5’ RNA cleavage product purified by Urea-PAGE, extracted from macerated gel using a 0.22 uM centrifugal filter (Corning, USA) as described previously7 (link) and ethanol precipitated. The cleavage product was resuspended in nuclease-free water and analysed by MALDI-ToF mass spectrometry using an Ultraflex III TOF-TOF instrument (Bruker Daltonik, Germany) in positive ion mode as described previously2 (link). Expected masses were calculated using the Mongo Oligo Mass Calculator v.2.06. The 5’ “6FAM” (6-Carboxyfluorescien)(IDT, USA) has a mass of 537.5 Da.
0.22 um centrifugal filter
Manufactured by Corning
Sourced in United States
The 0.22 μM centrifugal filter is a laboratory device used for the separation and concentration of macromolecules, such as proteins, nucleic acids, and other biological molecules, from complex mixtures. It utilizes a membrane with a pore size of 0.22 microns to selectively retain the desired molecules while allowing smaller components to pass through during the centrifugation process.
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Lab products found in correlation
2 protocols using 0.22 um centrifugal filter
1
XNAzyme-mediated RNA Cleavage Analysis
2
XNAzyme-mediated RNA Cleavage Analysis
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1 uM “Sub_Ebo” RNA was reacted with XNAzyme “FR6_1” under high magnesium selection conditions (20 h, 17 °C) and the 5’ RNA cleavage product purified by Urea-PAGE, extracted from macerated gel using a 0.22 uM centrifugal filter (Corning, USA) as described previously7 (link) and ethanol precipitated. The cleavage product was resuspended in nuclease-free water and analysed by MALDI-ToF mass spectrometry using an Ultraflex III TOF-TOF instrument (Bruker Daltonik, Germany) in positive ion mode as described previously2 (link). Expected masses were calculated using the Mongo Oligo Mass Calculator v.2.06. The 5’ “6FAM” (6-Carboxyfluorescien)(IDT, USA) has a mass of 537.5 Da.
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