A Total Extract Dot Blot Hybridization Procedure for mRNA Quantitation in Small Samples of Tissues or Cultured Cells
A Total Extract Dot Blot Hybridization Procedure for mRNA Quantitation in Small Samples of Tissues or Cultured Cells
- Analytical BioChemistry, 172(2), p.436-443, 1988 .
A simple method for the estimation of specific mRNA concentrations in small tissue samples (as little as 1 mg)or cultured cells (lower limit IO5 cells)is described. Guanidine hydrochloride extracts of whole cells or tissues are applied directly onto nitrocellulose and hybridized with the appropriate nick-translated probe. Loading according to DNA content allows expression ofthe result as concentration per cell. Hybridizing with a ribosomal RNA probe allows expression of results relative to rRNA and estimation of the RNA/DNA ratio in the sample. We describe the application of this procedure to the measurement of ceruloplasmin mRNA in tissues and cultured hepatocytes.
RECOMBINANT DNA TECHNOLOGY
MRNA
RNA: DNA: GENE EXPRESSION
A simple method for the estimation of specific mRNA concentrations in small tissue samples (as little as 1 mg)or cultured cells (lower limit IO5 cells)is described. Guanidine hydrochloride extracts of whole cells or tissues are applied directly onto nitrocellulose and hybridized with the appropriate nick-translated probe. Loading according to DNA content allows expression ofthe result as concentration per cell. Hybridizing with a ribosomal RNA probe allows expression of results relative to rRNA and estimation of the RNA/DNA ratio in the sample. We describe the application of this procedure to the measurement of ceruloplasmin mRNA in tissues and cultured hepatocytes.
RECOMBINANT DNA TECHNOLOGY
MRNA
RNA: DNA: GENE EXPRESSION