Hybridization blotting

Blotting Hybridization Techniques Allow Visualization of Specific Fragments... 

Oser, A., Roth, W.K., and Valet, G. (1988) Sensitive non-radioactive dot-blot hybridization using DNA probes labeled with chelate group substituted psoralen and quantitative detection by europium ion fluorescence. Nucleic Acids Res. 16, 1181-1196. 

B) RNA gel blot hybridization revealed massive enhancement of the PAP1 expression. Total RNA was isolated from 4-week-old papl-D and wildtype plants. 

Hybridization can be performed by merely spotting the sample to a membrane, where it is immobilized by baking and subsequently hybridized to a suitable probe. Sample application can be performed with commercially available manifolds that apply sample into multiple wells of the manifold and let sample migrate as spots or slots into the membrane hence the name dot blot or slot blot hybridization (W2). The sample wells are repeatedly washed prior to removing the membrane to bake or irradiate in order to fix the sample, which is then ready for hybridization with probe. 

Traditionally, Northern blot analysis of isolated total RNA or mRNA has been used to determine differences in levels of mRNA between and among different animal groups, tissues or time-points. Standard protocols for performing Northern blot hybridization are available (Sambrook and Russell, 2001). Radioisotopic and colorimetric methods are available for probe labeling. The advantage of Northern blot analysis, especially... 

Figure 4. Comparative detection of 4-chlorobiphenyl catabolic gene abundance by blot hybridization of pSSSO plasmid DNA probe to target DNA extracted from sediments. A B spatially separated sediments C control self hybridizations. (Units A B, ug total DNA C ng of probe DNA.)...

Gl. Gold, B., Radu, D., et al., Diagnosis of fragile X syndrome by Southern blot hybridization using a chemiluminescent probe A laboratory protocol. Mol. Diagn. 5(3), 169-178 (2000). 

Chan, W. T.-W., Fleming, K. A., and McGee, J. O D. (1985) Detection of subpicogram quantities of specific DNA sequences on blot hybridization with biotinylated probes Nucleic Acids Res. 13,8083-8091... 

Tsuda, H., Tani, Y, Hasegawa, T., and Fukutomi, T. 2001. Concordance in judgments among c-erbB-2 (HER2/neu) overexpression detected by two immunohistochemical tests and gene amplification detected by southern blot hybridization in breast carcinoma. Pathol. Internat. 51 26-32. 

Southern blot hybridization analysis proved that homologes of acyA and acyBl were almost perfectly conserved in the genomes of streptomycetes producing the... 

Pepin R, Lucas D, Lang R, Lee N, Liao M, Testa D (1990), Detection of picogram amounts of nucleic acid by dot blot hybridization, BioTechniques 8 628-632. 

Fig. 2D. Time course of degradation of calbindin-D28K mRNA in vitamin D-replete chick intestine following administration of cycloheximide. RNA was measured by dot blot hybridization to a calbindin-D2Rk cDNA probe.

Because of the sequence complexity of genomic DNA, isolation of false positive clones by blot hybridization is a significant problem when the stringency of the washing conditions is very low (wash temperature <42°C). 

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