RNA Fragmentation Reagent Kit
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- Electrophoresis
Supplier Data
Electrophoresis - RNA Fragmentation Reagent Kit (AB323459)
Size distribution of fragmented single-stranded RNA analyzed using Bioanalyzer 2100 Pico Chip series.
Figure 2. 1000 ng of purified single-stranded RNA fragmented in 1X Fragmentation Buffer and incubated at 94°C for times ranging from 0 minutes – 5 minutes and immediately stopped in 1X Stop Solution. Fragmented samples were diluted 1 : 10 in Nuclease-Free water and analyzed on the Bioanalyzer 2100, where relative size distribution of RNA fragments is graphed. Nucleotide size progressively decreases with fragmented samples treated with longer incubation times.
- Electrophoresis
Supplier Data
Electrophoresis - RNA Fragmentation Reagent Kit (AB323459)
Fragmentation of single-stranded RNA of a specified length on electrophoresis gel, showing varying fragment sizes controlled by incubation time of samples.
Figure 1. The fragment size of isolated RNA can be controlled by extending or shortening the incubation time of your samples at 94°C and immediately stopping the reaction with 1X Stop Solution. Figure 1 shows the fragmentation of single-stranded RNA samples that have been incubated at 94°C at times ranging from 0 minutes, 30 seconds – 4 minutes. Samples were precipitated and the length of fragments were visualized using nucleic acid electrophoresis gel. Samples that received longer incubation at 94°C show more fragmentation.
- Electrophoresis
Supplier Data
Electrophoresis - RNA Fragmentation Reagent Kit (AB323459)
Size distribution of fragmented eukaryotic mRNA analyzed using the Bioanalyzer 2100 Pico Chip series.
Figure 3. Isolated purified mRNA from eukaryotic cell pellet was fragmented with 1X Fragmentation Buffer and incubated at 94°C for 0 minutes, 30 seconds, and 1-4 minutes then immediately stopped with 1X Stop Solution. Samples were diluted 1 : 10 in Nuclease-Free water and analyzed using the Bioanalyzer 2100. The size distribution of mRNA fragments decreases with increasing incubation times.
Product details
RNA Fragmentation Reagent Kit (ab323459) is a reagent set used to fragment isolated RNA and purified mRNA for downstream processes including RNA-seq and next generation sequencing. The kit provides a simple, reliable and fast method for RNA library preparation.
Isolated purified RNA is incubated with divalent cations at 94°C to fragment RNA into shorter oligonucleotide strands. The fragment size of RNA can be controlled by extending or shortening the incubation time and stopping the reaction with Fragmentation Stop Solution.
Prepare 1 µg of isolated RNA samples in PCR Tubes with 10X Fragmentation Buffer and incubate in thermal cycler at 94°C for 1-5 minutes to fragment RNA. Add 10X Fragmentation Stop Solution to samples to stop the fragmentation reaction. Clean up fragmented RNA by performing an Ethanol precipitation. Fragmented samples may be qualified using gel electrophoresis to assess the yield and size distribution of fragmented RNA samples.
Performing a DNase-treatment on isolated RNA at sample prep is recommended for optimal results. Removal of buffer salts from fragmented RNA can be achieved by standard cold ethanol precipitation.
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