We evaluated the usefulness of PCR assays that target the internal transcribed spacer (ITS) region for identifying mycobacteria at the species level. The conservative and species-specific ITS sequences of 33 species of mycobacteria were analyzed in a multialignment analysis. One pair of panmycobacterial primers and seven pairs of mycobacterial species-specific primers were designed. All PCRs we...

Eggs of oviparous species must contain all components required for normal embryonic development. Among these, the vitamin riboflavin is deposited into egg white and yolk bound to the carrier protein, riboflavin-binding protein (ribBP). We have begun to investigate the mechanisms underlying ribBP transport pathways by molecular characterization of a relevant mutation in chicken. The aut...

Aim. To determine the influence of alternative alleles Ppd-D1 gene and phenotypic differences in duration need for vernalization their interaction on earing time grain yield recombinant lines bread winter wheat. Methods. Growing plants field vegetation site, PCR with specific primers Ppd-B1c genes, phenological observations, artificial vernalization, statistical analysis. Results. 136 F7 from c...

We report the development of a self-contained (homogeneous), single-tube assay for the genotyping of single-nucleotide polymorphisms (SNPs), which does not rely on fluorescent oligonucleotide probes. The method, which we call Tm-shift genotyping, combines allele-specific PCR with the discrimination between amplification products by their melting temperatures (Tm). Two distinct forward primers, ...

Major histocompatibility complex (MHC) class I chain-related gene B (MICB) encodes a ligand for activating NKG2D that expressed in natural killer cells, γδ T cells, and αβ CD8+ T cells, which is associated with autoimmune diseases, cancer, and infectious diseases. Here, we have established a system for genotyping MICB alleles using allele-specific primer extension (ASPE) on microarrays. Thirty-...

We describe here a quantitative real-time PCR assay for the detection of single-base-pair differences that does not require fluorescently labeled gene-specific probes or complicated primer combinations. Following PCR or RT-PCR of a gene segment that may contain allele-specific differences, 100 pg amplified product are used for a real-time PCR with allele-specific primers and SYBR Green. The use...

The detection of single nucleotide polymorphisms (SNPs) is becoming increasingly important in both the evaluation of candidate gene polymorphisms and fine mapping studies (4,7). Many techniques exist for the detection of SNPs within random sequences, such as single-strand conformation polymorphism (SSCP) (5,6), heteroduplex analysis and direct sequencing, but few techniques specifically identif...

Plasma membrane H+-ATPase is a major integral membrane protein with a role in various physiological processes including abiotic stress response. To study the effect of NaCl on the expression pattern of a gene encoding the plasma membrane H+-ATPase, an experiment was carried out in a completely random design with three replications. A pair of specific primers was designed based on the sequence o...