Formaldehyde-free RNA electrophoresis on denaturing agarose gels

Cat. No. AMR-N726-KIT

Features:

• Hood-free pouring and running - completely formaldehyde-free.
• Single-step RNA denaturation and staining.
• Eliminate long incubations and buffer recirculation.
• Sample preparation - denature and stain samples in a single step.
• Instant band visualization - low background and brilliant green bands without the need for ethidium bromide.
• Compatible with down-stream applications including Northern Blots.
• Glyoxol-free formulation.

• Formaldehyde-Free RNA Gel Solution, 10X
• Formaldehyde-Free RNA Running Buffer, 10X
• Formaldehyde-Free RNA Loading Buffer, 2X - bromophenol blue and xylene cyanol included as tracking dyes.

Comparison of Formaldehyde-Free RNA dye to ethidium bromide staining. Total RNA was extracted from K562 cells with Ribozol® RNA Extraction Reagent (N580). Samples were denatured in Formaldehyde-Free RNA Loading Buffer containing either ethidium bromide or Formaldehyde-Free RNA dye and incubated for 10 minutes at 65°C. The samples were applied to a 2% Formaldehyde-Free RNA Agarose Gel (Agarose I™, AMR-0710) and resolved for 1.5 hours at 5.1 V/cm. Image capture was performed with a Syngene GBox-HR Gel Doc System with a SYBR® Green filter.

Northern blot analysis of PGK1 mRNA resolved on Formaldehyde-Free RNA Gel. Data provided by Jeff Coller, Ph.D., Center for RNA Molecular Biology, CaseWestern ReserveUniversity, Cleveland, Ohio. Gel A: 20 μg of total yeast RNA was applied per lane to an agarose Formaldehyde-Free RNA Gel and resolved at 8V/cm. Bands were visualized on a UV transilluminator Gel B: Bands were transferred to Hybond N membrane and probed overnight with a radiolabelled oligonucleotide to PGK1 mRNA. Gel C: Membranes were stripped and reprobed over night with an oligonucleotide to SCR1 RNA loading control.
Lane 1: Untransformed control
Lane 2: High Copy PGK1 vector
Lane 3: Low Copy PGK1 vector
Lane 4: High Copy PGK1 vector
Lane5: Low Copy PGK1 vector