Gelatin as a blocking agent in Southern blot and chromosomal in situ hybridizations

1993 ◽  
Vol 9 (8) ◽  
pp. 261-262 ◽  
Author(s):  
S.C. Lakhotia ◽  
Abbay Sharma ◽  
Mousumi Mutsuddi ◽  
Madhu G. Tapadia
Keyword(s):  
Southern Blot ◽  
Blocking Agent ◽  
In Situ Hybridizations

scholarly journals Germline Transformation of Drosophila virilis With the Transposable Element mariner

Genetics ◽  
1996 ◽  
Vol 143 (1) ◽  
pp. 365-374 ◽  
Author(s):  
Allan R Lohe ◽  
Daniel L Hartl
Keyword(s):  
Transposable Element ◽  
Common Ancestor ◽  
Pcr Amplification ◽  
Drosophila Virilis ◽  
Cell Region ◽  
Diverse Species ◽  
Drosophila Mauritiana ◽  
Characteristic 2 ◽  
In Situ Hybridizations

Abstract An important goal in molecular genetics has been to identify a transposable element that might serve as an efficient transformation vector in diverse species of insects. The transposable element mariner occurs naturally in a wide variety of insects. Although virtually all mariner elements are nonfunctional, the Mosl element isolated from Drosophila mauritiana is functional. Mosl was injected into the pole-cell region of embryos of D. virilis, which last shared a common ancestor with D. mauritiana 40 million years ago. Mosl PCR fragments were detected in several pools of DNA from progeny of injected animals, and backcross lines were established. Because Go lines were pooled, possibly only one transformation event was actually obtained, yielding a minimum frequency of 4%. Mosl segregated in a Mendelian fashion, demonstrating chromosomal integration. The copy number increased by spontaneous mobilization. In situ hybridization confirmed multiple polymorphic locations of Mosl. Integration results in a characteristic 2-bp TA duplication. One Mosl element integrated into a tandem array of 370-bp repeats. Some copies may have integrated into heterochromatin, as evidenced by their ability to support PCR amplification despite absence of a signal in Southern and in situ hybridizations.

scholarly journals Persistence of chronic myelocytic leukemia despite deletion of rearranged bcr/c-abl sequences in blast crisis.

1986 ◽  
Vol 164 (5) ◽  
pp. 1389-1396 ◽  
Author(s):  
C R Bartram ◽  
J W Janssen ◽  
R Becher ◽  
A de Klein ◽  
G Grosveld
Keyword(s):  
In Situ Hybridization ◽  
T Cell ◽  
Southern Blot ◽  
Blast Crisis ◽  
Cell Lineage ◽  
Chronic Myelocytic Leukemia ◽  
Northern Blots ◽  
Myelocytic Leukemia ◽  
Ph Chromosome

We report on a Ph+ chronic myelocytic leukemia (CML) case, cytogenetically characterized by the occurrence of a second Philadelphia (Ph) chromosome in lymphoid blast crisis of T cell lineage. In situ hybridization analyses showed a deletion of translocated c-abl sequences, present on the Ph during chronic state, from both Ph in acute state. Moreover, Southern blot analyses of blastic cells exhibited a rearrangement within bcr, but a deletion of 5' bcr sequences, and Northern blots failed to detect the hybrid 8.5 kb bcr/c-abl transcript usually observed in Ph+ CML.

scholarly journals Methods for the analysis of large gene deletions and their application in some monogenic disorders

2009 ◽  
Vol 150 (50) ◽  
pp. 2258-2264 ◽  
Author(s):  
Péter Gergics ◽  
Judit Tőke ◽  
Ágnes Szilágyi ◽  
Ágnes Szappanos ◽  
Zoltán Kender ◽  
...  
Keyword(s):  
Southern Blot ◽  
Gene Deletions ◽  
Von Hippel Lindau ◽  
Monogenic Disorders ◽  
Large Gene

Számos monogénesen öröklődő kórképben a betegséget okozó gén teljes vagy részleges deletiója, illetve kópiaszámának megváltozása patogenetikai tényezőként jön számításba. A direkt DNS-szekvenálás nem alkalmas a gén nagy deletiójának, illetve kópiaszám-változásának kimutatására. Az összefoglalóban a szerzők áttekintik a nagy géndeletio vizsgálómódszereit, és két, monogénesen öröklődő betegségben végzett saját vizsgálataik példáján keresztül bemutatják a módszerek gyakorlati alkalmazásának lehetőségeit. Vázolják a géndeletio-vizsgálat hagyományos (kromoszóma-sávtechnika, Southern-blot, fluoreszcens in situ hibridizáció) és polimeráz láncreakcióra alapozott módszereit (denaturáló nagy felbontóképességű folyadékkromatográfia, kvantitatív valós idejű polimeráz láncreakció, mikroszatellitamarker-analízis, multiplex amplifikálhatópróba-hibridizáció, multiplex ligatióspróba-analízis), valamint a technikai és informatikai haladás legújabb vívmányait (komparatív genomhibridizálás, „array” analízis). Saját vizsgálataikban von Hippel–Lindau-szindrómában szenvedő betegekben kvantitatív valós idejű polimeráz láncreakció és multiplex ligatióspróba-amplifikálás alkalmazásával bemutatják a VHL, illetve congenitalis adrenalis hyperplasiás betegekben a CYP21A2 géndeletio-vizsgálat eredményeit és ezek klinikai jelentőségét.

Waldenström macroglobulinemia neoplastic cells lack immunoglobulin heavy chain locus translocations but have frequent 6q deletions

Blood ◽  
2002 ◽  
Vol 100 (8) ◽  
pp. 2996-3001 ◽  
Author(s):  
Roelandt F. J. Schop ◽  
W. Michael Kuehl ◽  
Scott A. Van Wier ◽  
Gregory J. Ahmann ◽  
Tammy Price-Troska ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Tumor Cells ◽  
Southern Blot ◽  
Heavy Chain ◽  
Genomic Stability ◽  
Immunoglobulin M ◽  
Lymphoplasmacytic Lymphoma ◽  
Interphase Fish

Lymphoplasmacytic lymphoma (LPL) is characterized by t(9;14)(p13;q32) in 50% of patients who lack paraproteinemia. Waldenström macroglobulinemia (WM), which has an immunoglobulin M (IgM) paraproteinemia, is classified as an LPL. Rare reports have suggested that WM sometimes is associated with 14q23 translocations, deletions of 6q, and t(11;18)(q21;q21). We tested for these abnormalities in the clonal cells of WM patients. We selected patients with clinicopathologic diagnosis of WM (all had IgM levels greater than 1.5 g/dL). Southern blot assay was used to detect legitimate and illegitimate IgH switch rearrangements. In addition to conventional cytogenetic (CC) and multicolor metaphase fluorescence in situ hybridization (M-FISH) analyses, we used interphase FISH to screen for t(9;14)(p13;q32) and other IgH translocations, t(11;18)(q21;q21), and 6q21 deletions. Genomic stability was also assessed using chromosome enumeration probes for chromosomes 7, 9, 11, 12, 15, and 17 in 15 patients. There was no evidence of either legitimate or illegitimate IgH rearrangements by Southern blot assay (n = 12). CC (n = 37), M-FISH (n = 5), and interphase FISH (n = 42) failed to identify IgH or t(11;18) translocations. Although tumor cells from most patients were diploid for the chromosomes studied, deletions of 6q21 were observed in 42% of patients. In contrast to LPL tumors that are not associated with paraproteinemia and that have frequent t(9;14)(p13;q32) translocations, IgH translocations are not found in WM, a form of LPL tumor distinguished by IgM paraproteinemia. However, WM tumor cells, which appear to be diploid or near diploid, often have deletions of 6q21.

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