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Genes - University of Arizona | Ecology and Evolutionary Biology
Genes - University of Arizona | Ecology and Evolutionary Biology

... DNA Technology The following are some of the most important molecular methods we will be using in this course. They will be used, among other things, for ...
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... •DNA fragments can be synthesized by highly automated machines. •Nucleotide sequences in a stretch of DNA can also be determined using automated machines. If the DNA is long, it is first cut into smaller fragments by restriction enzymes, and the fragments are sequenced separately. •Sequences are sto ...
Human Genome Project and Sequencing
Human Genome Project and Sequencing

... Humans are 99.9% identical. Total number of genes ~ 30,000. This doesn’t match the number of proteins (over 100,000) so each gene must be able to code for more than one protein. Over 50% of genes have unknown ...
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File - Mrs. LeCompte
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... Foreign DNA can be incorporated into a cloning vector (plasmid, phage, YAC or other) if both foreign DNA and vector are cleaved with the same restriction endonuclease. The pieces of DNA will anneal, and then can be ligated using DNA ligases. The desired fragment can be separated from others using g ...
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Biology_Ch._14
Biology_Ch._14

Document
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Comparative genomic hybridization



Comparative genomic hybridization is a molecular cytogenetic method for analysing copy number variations (CNVs) relative to ploidy level in the DNA of a test sample compared to a reference sample, without the need for culturing cells. The aim of this technique is to quickly and efficiently compare two genomic DNA samples arising from two sources, which are most often closely related, because it is suspected that they contain differences in terms of either gains or losses of either whole chromosomes or subchromosomal regions (a portion of a whole chromosome). This technique was originally developed for the evaluation of the differences between the chromosomal complements of solid tumor and normal tissue, and has an improved resoIution of 5-10 megabases compared to the more traditional cytogenetic analysis techniques of giemsa banding and fluorescence in situ hybridization (FISH) which are limited by the resolution of the microscope utilized.This is achieved through the use of competitive fluorescence in situ hybridization. In short, this involves the isolation of DNA from the two sources to be compared, most commonly a test and reference source, independent labelling of each DNA sample with a different fluorophores (fluorescent molecules) of different colours (usually red and green), denaturation of the DNA so that it is single stranded, and the hybridization of the two resultant samples in a 1:1 ratio to a normal metaphase spread of chromosomes, to which the labelled DNA samples will bind at their locus of origin. Using a fluorescence microscope and computer software, the differentially coloured fluorescent signals are then compared along the length of each chromosome for identification of chromosomal differences between the two sources. A higher intensity of the test sample colour in a specific region of a chromosome indicates the gain of material of that region in the corresponding source sample, while a higher intensity of the reference sample colour indicates the loss of material in the test sample in that specific region. A neutral colour (yellow when the fluorophore labels are red and green) indicates no difference between the two samples in that location.CGH is only able to detect unbalanced chromosomal abnormalities. This is because balanced chromosomal abnormalities such as reciprocal translocations, inversions or ring chromosomes do not affect copy number, which is what is detected by CGH technologies. CGH does, however, allow for the exploration of all 46 human chromosomes in single test and the discovery of deletions and duplications, even on the microscopic scale which may lead to the identification of candidate genes to be further explored by other cytological techniques.Through the use of DNA microarrays in conjunction with CGH techniques, the more specific form of array CGH (aCGH) has been developed, allowing for a locus-by-locus measure of CNV with increased resolution as low as 100 kilobases. This improved technique allows for the aetiology of known and unknown conditions to be discovered.
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