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Successful investigation of common diseases requires advances in our understanding of the organization of the genome. Linkage disequilibrium provides a theoretical basis for performing candidate gene or whole-genome association studies to analyze complex disease. However, to constructively interrogate SNPs for these studies, technologies with sufficient throughput and sensitivity are required. A plethora of suitable and reliable methods have been developed, each of which has its own unique advantage. The characteristics of the most promising genotyping and polymorphism scanning technologies are presented. These technologies are examined both in the context of complex disease investigation and in their capacity to face the unique physical and molecular challenges (allele amplification, loss of heterozygosity and stromal contamination) of solid tumor research. 相似文献
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Universal DNA array detection of small insertions and deletions in BRCA1 and BRCA2 总被引:19,自引:0,他引:19
Array-based mutation detection methodology typically relies on direct hybridization of the fluorescently labeled query sequence to surface-bound oligonucleotide probes. These probes contain either small sequence variations or perfect-match sequence. The intensity of fluorescence bound to each oligonucleotide probe is intended to reveal which sequence is perfectly complementary to the query sequence. However, these approaches have not always been successful, especially for detection of small frameshift mutations. Here we describe a multiplex assay to detect small insertions and deletions by using a modified PCR to evenly amplify each amplicon (PCR/PCR), followed by ligase detection reaction (LDR). Mutations were identified by screening reaction products with a universal DNA microarray, which uncouples mutation detection from array hybridization and provides for high sensitivity. Using the three BRCA1 and BRCA2 founder mutations in the Ashkenazi Jewish population (BRCA1 185delAG; BRCA1 5382insC; BRCA2 6174delT) as a model system, the assay readily detected these mutations in multiplexed reactions. Our results demonstrate that universal microarray analysis of PCR/PCR/LDR products permits rapid identification of small insertion and deletion mutations in the context of both clinical diagnosis and population studies. 相似文献
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低温对植物叶片中超氧物歧化酶、过氧化氢酶和过氧化氢水平的影响 总被引:10,自引:0,他引:10
番茄和鸡蛋果叶片中可提取的SOD活性不受低温的影响。在电泳谱带上SOD主同工酶带被氰化物而不被低温抑制,次同工酶带在低温下不稳定,且活性很低,它的变化不影响总的SOD活性。一些冷敏感植物叶片中CAT活性被低温抑制,而H_2O_3水平在低温下稳定或有增加,这可能使毒性更强的羟基离子(OH·)易于形成。 相似文献
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Adam Doern Xianjun Cao Arlene Sereno Christopher L. Reyes Angelina Altshuler Flora Huang Cathy Hession Albert Flavier Michael Favis Hon Tran Eric Ailor Melissa Levesque Tracey Murphy Lisa Berquist Susan Tamraz Tracey Snipas Ellen Garber William S. Shestowsky Rachel Rennard Christilyn P. Graff Xiufeng Wu William Snyder Lindsay Cole David Gregson Michael Shields Steffan N. Ho Mitchell E. Reff Scott M. Glaser Jianying Dong Stephen J. Demarest Kandasamy Hariharan 《The Journal of biological chemistry》2009,284(15):10254-10267
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Wingfield BD; Grant WS; Wolfaardt JF; Wingfield MJ 《Molecular biology and evolution》1994,11(3):376-383
The genus Ceratocystis sensu stricto includes important fungal pathogens of
woody and herbaceous plants. This genus is distinguished from species in
Ceratocystis sensu lato by the presence of Chalara anamorphs. Ascospore
shape has been used extensively in delineating Ceratocystis taxa, which
show a large variety of ascospore shapes. Sequence analysis of one region
of he 18S ribosomal RNA subunit and two regions of the 28S ribosomal RNA
subunit showed that there was a majority of multiple substitutions at
nucleotide sites and that there was a low transition/transversion ratio, T
= 0.72. Both of these results suggest that these are well established, old
species. Ascospore morphology, for the most part, was not congruent with
the molecular phylogeny, and the use of morphological characters may be
misleading in the taxonomy of these species.
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BD Llewellyn 《Biotechnic & histochemistry》2013,88(4):159-177
The hematoxylin and eosin stain is the most common method used in anatomic pathology, yet it is a method about which technologists ask numerous questions. Hematoxylin is a natural dye obtained from a tree originally found in Central America, and is easily converted into the dye hematein. This dye forms coordination compounds with mordant metals, such as aluminum, and the resulting lake attaches to cell nuclei. Regressive formulations contain a higher concentration of dye than progressive formulations and may also contain a lower concentration of mordant. The presence of an acid increases the life of the solution and in progressive solutions may also affect selectivity of staining. An appendix lists more than 60 hemalum formulations and the ratio of dye to mordant for each. 相似文献