60 results about "Bac clone" patented technology

The invention discloses a reagent for detecting a copy number of an EGFR gene and the ploidy of a chromosome 7. The reagent comprises a fluorescence probe RP11-339F13 used for detecting the copy number of the EGFR gene and a fluorescence probe RP11-144H20 used for detecting the ploidy of the chromosome 7, wherein the RP11-339F13 is cloned by BAC, which covers the whole EGFR gene, and marked with rhodamine fluorescein; and RP11-144H20 is cloned by specific BAC of the centromere of the chromosome 7 and marked with fluorescein isothiocyanate. The reagent can quickly, simply and conveniently detect the copy number of the EGFR gene and the ploidy of the chromosome 7, contributes to the judgment of medicament susceptibility and prognostic judgment, and has great significance for the guidance of use of molecular targeted medicament tyrosine kinase inhibitors (TKIs) of tumors, such as non-small cell lung cancer and the like.

A single base change in the Bn-FAE1.1 gene in the A genome and a two-base deletion in the Bn-FAE1.2 gene in the C genome produce the nearly zero content of erucic acid observed in canola. A BAC clone anchoring Bn-FAE1.1 from a B. rapa BAC library and a BAC clone anchoring Bn-FAE1.2 from a B. oleracea BAC library were used in this research. After sequencing the gene flanking regions, it was found that the dissimilarity of the flanking sequences of these two FAE1 homologs facilitated the design of genome specific primers that could amplify the corresponding genome in allotetraploid B. napus. The two-base deletion in the C genome gene was detected as a sequence characterized sequence region (SCAR) marker. To increase the throughput, one genome specific primer was labeled with four fluorescence dyes and combined with 20 different primers to produce PCR products with different fragment sizes. Eventually, a super pool of 80 samples was detected simultaneously, making it possible to analyze over half a million of samples per day using a medium capacity ABI 3100 Genetic Analyzer. This dramatically reduces the cost of marker detection. The single base change in the Bn-FAE1.1 gene was detected as single nucleotide polymorphic (SNP) marker with an ABI SNaPshot kit. A multiplexing primer set was designed by adding a polyT to the 5′ primer end to increase SNP detection throughput through sample pooling. These multiplexed high throughput molecular markers have been successfully implemented in our canola / rapeseed breeding programs.