Hybridization-based genotyping

Using double-stranded PGR amplicons as target, ohgomer probes querying the sense as well as the antisense strand are on the microarray. Having only two possible reactions in either strand, study of cytosine methylation is less complex than traditional genotyping with potential incorporation of four different bases. Compared to our hybridization-based epigenetic arrays. 

Mycobacteria of the Mycobacterium avium complex are implicated in disseminated bacterial infections in AIDS patients. RFLP studies followed by hybridization with radiolabeled probe specific for an insertion sequence in M. avium (IS 1311) have been useful for typing M. avium stains (R2). A variety of molecular techniques are available for the diagnosis of Chlamydia trachomatis infection. In addition to PCR, a method based on the ligase chain reaction has also been found to be sensitive to the detection of C. trachomatis infection in urine specimens collected from male and female subjects (VI). The differentiation between low-risk genotypes of human papilloma virus (HPV 6 or 11) from genotypes of high... 

Standards based on this approach, and derived feed formulas, are shown in the following tables. Producers using modem hybrids instead of traditional genotypes are advised instead to use the NRC (1994) values or the values suggested by the breeding company as the basis for dietary standards. 

Schutz E, von Ahsen N, Oellerich M. Genotyping of eight thiopurine methyltransferase mutations three-color multiplexing, two-color/shared anchor, and fluorescence-quenching hybridization probe assays based on thermodynamic nearest-neighbor probe design. Clin Chem 2000 46 1728-1737. 

Fig. 2. Mutagenesis strategy. A neo gene replaced one-third of the ORF and was used as a positive selectable marker. The HSV thymidine gene (HSV-ffc) was used for negative selection. The vector was linearized with Xbal digestion. The targeted events were screened by Southern blot. Two Xbal sites are in the outside of the targeting vector (the 3 -end one is just at the end of the vector, which cannot be digested by Xbal if the vector is randomly inserted). An outside probe is located at the 5 -end. The genotype of the mice can be easily identified based on the size of the hybridized bands, 6.7 kb (mutant) vs 6 kb (wild-type).

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