ELECTROPHORESIS HEADER-2

Gel Electrophoresis of DNA and RNA

Methylation & Uracil Interference Assays

By National Diagnostics | August 22, 2011 | Comments Off on Methylation & Uracil Interference Assays

Both the methylation and uracil interference assay analytical methods are presented in parallel because they assay for similar information based on similar reactions. In both cases, the DNA is analyzed for nucleotides which are important…

Analysis of DNA/Protein Interactions

By National Diagnostics | August 19, 2011 | Comments Off on Analysis of DNA/Protein Interactions

The binding of proteins to specific DNA sites is an important mechanism of cellular regulation. Numerous techniques have been developed to analyze the interactions of regulatory proteins with DNA. Three techniques are presented here which…

Primer Extension

By National Diagnostics | August 19, 2011 | Comments Off on Primer Extension

Primer extension is another technique used to analyze RNA structure and expression. In this method, an oligonucleotide primer is annealed to RNA and extended to a cDNA copy by reverse transcriptase in the presence of…

Ribonuclease Protection

By National Diagnostics | August 19, 2011 | Comments Off on Ribonuclease Protection

Ribonuclease protection is a procedure that uses uniformly labeled RNA probes to analyze sample RNA. In this case, probes are chosen to fall entirely within the coding region, so they are only digested if no…

S1 Mapping

By National Diagnostics | August 19, 2011 | Comments Off on S1 Mapping

Nuclease S1 will digest only ssDNA or ssRNA. If a duplex of DNA and/or RNA strands have single-stranded overhangs or unhybridized internal loops, these will be digested away. The remaining intact nucleic acid fragments represent…

RNA Mapping

By National Diagnostics | August 19, 2011 | Comments Off on RNA Mapping

In studies of transcriptional regulation, it is often necessary to determine the structure and/or amount of a given RNA species. Several techniques have been developed making use of the fact that some nucleases will only…

Differential Display

By National Diagnostics | August 17, 2011 | Comments Off on Differential Display

Differential display is a powerful technique for detecting and quantitating changes in gene expression patterns between differently treated cells. Fragments of those genes which are induced or suppressed can be identified and isolated for further…

Automated Sequencers

By National Diagnostics | August 17, 2011 | Comments Off on Automated Sequencers

Automated sequencing systems make use of fluorescent dye labeling, in combination with laser scanning and computerized data acquisition and processing to carry out the electrophoresis of up to 96 sequencing reactions on a single gel,…

Pouring Sequencing Gels

By National Diagnostics | August 17, 2011 | Comments Off on Pouring Sequencing Gels

Denaturing PAGE gels for DNA sequencing generally employ 6-8 M urea as their denaturant and TBE as their buffer system. They are poured as described in the section on denaturing PAGE of DNA and RNA.…

Sanger Sequencing

By National Diagnostics | August 17, 2011 | Comments Off on Sanger Sequencing

In Sanger dideoxy terminator sequencing, the sample DNA is used as a template for a DNA polymerase. Four polymerase reactions are carried out involving enzyme, primer, and sample DNA, along with dNTPs. Each reaction also…