Anchoring enzyme digestion of cDNA
If the previous step in the series of analytical techniques for gene expression, SAGE cDNA, was stored at -20°C, then this experiment was thawed slowly on ice.
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Recovery of DNA from low melting point agarose gels (digestion with agarase)
DNA can be recovered from low melting point agarose gels by agarase digestion (Bumeister and lehrach 1989). In this method, the agarase enzyme hydrolyzes the ag
Phosphorylation of 5' prominence DNA molecules by exchange reaction
The T4 phage polynucleotide kinase-catalyzed exchange reaction (Van de Sande et al. 1973; Berkner and Folk 1977) is a rapid method for labeling 5' end DNA. Unli
Long-range PCR
Under standard PCR reaction conditions, PCR amplification of DNA fragments is generally 1-2 kb in length, which is sufficient for many routine DNA molecular man
Toxicological effects of environmental toxins detected by cDNA macroarrays and gene expression profiles
This chapter describes how to prepare and use cDNA macroarrays to detect the effects of environmental toxins on gene expression, and describes the materials and
Comparative Molecular Physiological Genomics - Heterozygous hybridization experiments with cDNA arrays
In recent years, DNA microarrays have been recognized as a standard method for molecular biology research. Especially in biomedical research, microarrays of com
Phosphorylation of DNA molecules containing a 5' prominent hydroxyl terminus
This protocol removes the 5' phosphate of a nucleic acid by phosphatase, and then re-adds the phosphate to the nucleic acid in a radiolabeled form catalyzed by
Single-cell mRNA differential display assay
Several methods exist to show changes in the expression levels of unknown genes in tissue samples in response to physiological stimuli. However, the morphologic
Radiolabeling of minus cDNA probes by random oligonucleotide extension method
The synthesis of cDNA in this protocol was carried out in four saturating concentrations of dNTP and one trace of radiolabeled dNTP. After deduction of hybridiz