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| Untersuchte Arbeit: Seite: 34, Zeilen: 12-23, 25-28 |
Quelle: Katzaki 2009 Seite(n): 71, Zeilen: left col. 18-34, 43-48 |
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| Multiplex Ligation-dependant Probe Amplification (MLPA) analysis
We used a distinct commercially available MLPA kit, the SALSA P036D subtelomeric primer set (MRC-Holland, Amsterdam, The Netherlands). This kit contains oligonucleotide primer sets specific for the amplification of selected loci in the subtelomeric regions of all chromosome arms, except for the acrocentric chromosomes 13, 14, 15, 21 and 22 that effectively lack a short arm. For the latter, the manufacturer has included in this kit primer sets specific for loci adjacent to the centromere in the long arm of the acrocentric chromosomes, referred to as the ‘acrocentric’ primer. This kit was previously validated in other laboratories (data not shown) on series of patients with known subtelomeric ultra conserved regions (UCRs) [29, 30]. The target loci of this kit represent known functional genes or protein coding sequences. Each experiment was carried out according to the manufacturer’s instructions. Fluorescent in situ hybridization (FISH) analysis Chromosomal preparations for the analysis were obtained according to standard techniques. FISH was performed with TelVision 9p and 17p probes (Vysis). Each experiment was carried out according to the manufacturer’s instructions. [29] Ahn J W, Ogilvie C M, Welch A, Thomas H, Madula R, Hills A, Donaghue C, Mann K. (2007) Detection of subtelomere imbalance using MLPA: validation, development of an analysis protocol, and application in a diagnostic centre. BMC Med Genet 8:9 [30] Rooms L, Reyniers E, van Luijk R, Scheers S, Wauters J, Ceulemans B, Van Den Ende J, Van Bever Y, Kooy R F. (2004) Subtelomeric deletions detected in patients with idiopathic mental retardation using multiplex ligation-dependent probe amplification (MLPA). Hum Mutat 23:17-21 |
MATERIAL AND METHODS
Multiplex Ligation-dependant Probe Amplification (MLPA) analysis We used a distinct commercially available MLPA kit, the SALSA P036D subtelomeric primer set (MRC-Holland, Amsterdam, The Netherlands). This kit contains oligonucleotide primer sets specific for the amplification of selected loci in the subtelomeric regions of all chromosome arms, except for the acrocentric chromosomes 13, 14, 15, 21 and 22 that effectively lack a short or p-arm. For the latter the manufacturer has included in this kit primer sets specific for loci adjacent to the centromere in the long arm of the acrocentric chromosomes, referred to as the ‘acrocentric’ primer. This kit was previously validated in other laboratories (data not shown) on series of patients with known subtelomeric UCRs [Ahn J et al. 2007, Kirchhoff et al. 2005, Rooms et al. 2004]. The target loci of this kit represent known functional genes or protein coding sequences. [...] FISH analysis Chromosomal preparations for the analysis were obtained according to standard techniques. Fluorescent in situ hybridization (FISH) was performed with TelVision 9p and 17p probes (Vysis). Each experiment was carried out according to the manufacturer’s instructions. |
Nothing has been marked as a citation. The original text is part of an article which is included in Katzaki's thesis. Mmu was not one of the coauthors of this article. The references for Ahn J et al. 2007, Kirchhoff et al. 2005, Rooms et al. 2004 are all missing in Katzaki (2009). |
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