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DNA Isolation from small tissue samples using salt and spermine
DNA Isolation from small tissue samples using salt and spermine

... and 0.4 ml of cold absolute ethanol. The DNA is allowed to precipitate either at -20°C overnight or at -80°C for 30 min. Centrifuge the samples for 15 minutes at 4°C at 17,000 g and wash the pellet twice in 70% ethanol by suspension and recentrifugation. The DNA pellet is dissolved in 1 ml of TE buf ...
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Biotechnology - Genetic Engineering

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Fruitful DNA Extraction
Fruitful DNA Extraction

... very similar genotypes while more distantly related organisms will have DNA sequences that have fewer base pairs in common. In this lab, we will be extracting DNA from kiwi fruit. Extracting DNA from an organism is the first step in determining its genotype. Typically, scientists will look at the se ...
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Chapter 12 DNA and RNA - Northwestern High School

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Title: Ready, Set, Clone! Authors: Kowalski, Kathiann M. Source

... Eppendorf tube. They add proteins called "restriction enzymes" to cut out the specific DNA section they want. They also add "primers/' which contain the A, C, G, and T nucleotides. The unwound DNA strands will need the primers to make the other half of the DNA molecule's helix or ladder. To help the ...
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... DNA FINGERPRINTING WITH PCR uses PCR to analyze highly variable microsatellite or minisatellite [aka VNTR (variable numbers of tandem repeats)] loci to determine DNA identity (as in forensic blood tests) or to determine parentage of an individual. Minisatellite sites are highly polymorphic* regions ...
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TE content correlates positively with genome size

< 1 ... 164 165 166 167 168 169 170 171 172 ... 222 >

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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