Enhanced sensitivity RNA gel loading buffer that enables efficient RNA separation on native gels
Author(s) -
Keqin Gregg,
Wenli Zhou,
Wan Ji,
Sara K. Davis
Publication year - 2004
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/04362pf01
Subject(s) - rna , agarose , ethidium bromide , rna extraction , agarose gel electrophoresis , nuclease protection assay , gel electrophoresis , microbiology and biotechnology , gene expression , chemistry , chromatography , dna microarray , biology , dna , gene , rna dependent rna polymerase , biochemistry
RNA gel analysis is essential for quality assessment of RNA preparations for subsequent analysis such as microarrays and real-time PCRs. The routinely used standard electrophoresis of RNA through formaldehyde-containing agarose gels is not only labor-intensive and time-consuming, but also involves sizeable quantities of hazardous materials. Above all, it is not sensitive, requiring more than 1 microgram of RNA for the assay. Current gene expression profiling with microarrays and real-time PCR often involves limiting amounts of RNA. It is therefore important to have a more sensitive way to analyze RNA. Here we report an improved ethidium bromide-based RNA gel analysis system with our Superload buffer that increases sensitivity to 12.5 ng of total RNA and allows RNA analysis on a regular native Tris-acetate EDTA (TAE) agarose gel.
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