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New insights into the pathogenic mechanisms associated with CNVs: duplication of 17p13.3, mirror effect in 16p11.2 and recessive phenotype in 22q11.22

von Dott. Mafalda Mucciolo

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Statistik und Sichtungsnachweis dieser Seite findet sich am Artikelende
[1.] Mmu/Fragment 008 01 - Diskussion
Zuletzt bearbeitet: 2014-11-18 19:11:53 Singulus
Fragment, Gesichtet, KomplettPlagiat, Mmu, SMWFragment, Schutzlevel sysop, Shinawi and Cheung 2008

Typus
KomplettPlagiat
Bearbeiter
Graf Isolan
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Untersuchte Arbeit:
Seite: 8, Zeilen: 1-18
Quelle: Shinawi and Cheung 2008
Seite(n): 761-762, 763, Zeilen: 761:right col. 5-8.11 - 762:left col. 1.4-10; Figure 2: legend 6-7; 763:left col. 6-11, right col. 10-12
The higher resolution and throughput with possibilities for automation, robustness, simplicity, high reproducibility and precise mapping of aberrations are the most significant advantages of aCGH over cytogenetic methods. In addition, there is no need for cell culture, making the turn around time shorter than in cytogenetic methods. As with other clinical diagnostic methods, there are limitations in aCGH technology. aCGH is not able to identify balanced rearrangements such as translocations and inversions and low-level mosaicism for unbalanced numeric or structural rearrangements.

1.2 Array – CGH Methodologies

In aCGH, equal amounts of labelled genomic DNA from a test and a reference sample are co-hybridized to an array containing the DNA targets. Genomic DNA of the patient and control are differentially labelled with Cyanine 3 (Cy3) and Cyanine 5 (Cy5). The slides are scanned into image files using a microarray scanner. The spot intensities are measured and the image files are quantified using feature extraction software, and text file outputs from the quantitative analyses are imported into software programs for copy number analysis (Fig.2) (Cheung 2005, Lu 2007). The resulting ratio of the fluorescence intensities is proportional to the ratio of the copy numbers of DNA sequences in the test and reference genomes.


17. Cheung S.W. et al. (2005) Development and validation of a CGH microarray for clinical cytogenetic diagnosis. Genet. Med. 7, 422–432

43. Lu, X. et al. (2007) Clinical implementation of chromosomal microarray analysis: summary of 2513 postnatal cases. PLoS ONE 2, e327

[Page 761]

aCGH methodology

In aCGH, equal amounts of labeled genomic DNA from a test and a reference sample are cohybridized to an array containing the DNA targets. [...] Genomic DNA of the patient and control are differen-

[Page 762]

tially labeled with Cyanine 3 (Cy3) and Cyanine 5 (Cy5) (Fig. 2a). [...] The spot intensities are measured (Fig. 2c) and the image files are quantified using feature extraction software, and text file outputs from the quantitative analyses are imported into software programs for copy number analysis (Fig. 2d) [4,9]. The resulting ratio of the fluorescence intensities is proportional to the ratio of the copy numbers of DNA sequences in the test and reference genomes.


FIGURE 2

[...] (b) The slides are scanned into image files using a specific microarray scanner.

[Page 763]

The advantages and limitations of diagnostic aCGH

The higher resolution and throughput with possibilities for automation, robustness, simplicity, high reproducibility and precise mapping of aberrations are the most significant advantages of aCGH over cytogenetic methods. In addition, there is no need for cell culture, making the turn around time shorter than in cytogenetic methods. [...]

As with other clinical diagnostic methods, there are limitations in aCGH technology. aCGH is not able to identify balanced rearrangements such as translocations and inversions.


4 Cheung, S.W. et al. (2005) Development and validation of a CGH microarray for clinical cytogenetic diagnosis. Genet. Med. 7, 422–432

9 Lu, X. et al. (2007) Clinical implementation of chromosomal microarray analysis: summary of 2513 postnatal cases. PLoS ONE 2, e327

Anmerkungen

Nothing has been marked as a citation. The source is not given.

Sichter
(Graf Isolan), SleepyHollow02

[2.] Mmu/Fragment 008 19 - Diskussion
Zuletzt bearbeitet: 2014-12-07 21:49:43 Singulus
Fragment, Gesichtet, KomplettPlagiat, Mmu, Papa 2010, SMWFragment, Schutzlevel sysop

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Bearbeiter
Graf Isolan
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Untersuchte Arbeit:
Seite: 8, Zeilen: 9, 19-30
Quelle: Papa 2010
Seite(n): 13, Zeilen: 10-23
1.2 Array – CGH Methodologies

[...]

Two major types of array targets are currently being utilized. Initially, bacterial artificial chromosomes (BACs) were the array target of choice (Pinkel 1998). However, now oligonucleotide arrays have been adopted due to the increased genome coverage they afford. The design of both array types was made possible by the availability of the complete map and sequence of the human genome. The BAC arrays may contain DNA isolated from large insert clones that range in size from 150–200 kb, spotted directly onto the array or may employ the spotting of PCR products amplified from the BAC clones (Ylstra 2006). These arrays are generally very sensitive and results can be directly validated with FISH using the BAC DNA as a probe. However, production of BAC DNA is labor-intensive, and the resolution is limited to 50–100 kb, even on a whole genome tiling path array (Ishkanian 2004). Oligonucleotide arrays offer a flexible format with the potential for very high [resolution and customization.]


30. Ishkanian A.S. et al. 2004. A tiling resolution DNA microarray with complete coverage of the human genome. Nat. Genet. 36: 299–303.

60. Pinkel D. et al. 1998. High resolution analysis of DNA copy number variation using comparative genomic hybridization to microarrays. Nat. Genet. 20: 207–211.

87. Ylstra B. et al. 2006. BAC to the future! Or oligonucleotides: A perspective for micro array comparative genomic hybridization (array CGH). Nucleic Acids Res. 34: 445–450.

1.2 Array – CGH Methodologies

Two major types of array targets are currently being utilized. Initially, bacterial artificial chromosomes (BACs) were the array target of choice. [6] However, more recently, oligonucleotide arrays have been adopted due to the increased genome coverage they afford. The design of both array types was made possible by the availability of the complete map and sequence of the human genome. The BAC arrays may contain DNA isolated from large insert clones that range in size from 150–200 Kb, spotted directly onto the array or may employ the spotting of PCR products amplified from the BAC clones. [8] These arrays are generally very sensitive and results can be directly validated with FISH using the BAC DNA as a probe. However, production of BAC DNA is labor-intensive and the resolution is limited to 50–100 Kb, even on a whole genome tiling path array. [9] Oligonucleotide arrays offer a flexible format with the potential for very high resolution and customization.


6. Pinkel, D., et al., High resolution analysis of DNA copy number variation using comparative genomic hybridization to microarrays. Nat Genet, 1998. 20(2): p. 207-11.

8. Ylstra, B., et al., BAC to the future! or oligonucleotides: a perspective for micro array comparative genomic hybridization (array CGH). Nucleic Acids Res, 2006. 34(2): p. 445-50.

9. Ishkanian, A.S., et al., A tiling resolution DNA microarray with complete coverage of the human genome. Nat Genet, 2004. 36(3): p. 299-303.

Anmerkungen

Nearly identical with identical references. Nothing has been marked as a citation.

Sichter
(Graf Isolan), SleepyHollow02


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