scholarly journals Polymerase chain reaction-based suppression of repetitive sequences in whole chromosome painting probes for FISH

2005 ◽  
Vol 13 (2) ◽  
pp. 27-32 ◽  
Author(s):  
Lawrence C. Dugan ◽  
Melissa S. Pattee ◽  
Jennifer Williams ◽  
Mike y ◽  
Karen Sorensen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chromosome Painting ◽  
Repetitive Sequences ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Painting Probes ◽  
Whole Chromosome Painting

scholarly journals A method for mapping germ line sequences in Tetrahymena thermophila using the polymerase chain reaction.

Genetics ◽  
1994 ◽  
Vol 137 (1) ◽  
pp. 95-106 ◽  
Author(s):  
D Cassidy-Hanley ◽  
M C Yao ◽  
P J Bruns
Keyword(s):  
Polymerase Chain Reaction ◽  
Dna Sequences ◽  
Tetrahymena Thermophila ◽  
Germ Line ◽  
Repetitive Sequences ◽  
Mapping Technique ◽  
Ciliated Protozoan ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Template Dna

Abstract A method for mapping DNA sequences to specific germinal chromosomes in the ciliated protozoan Tetrahymena thermophila has been developed. This mapping technique (PCR mapping) utilizes the polymerase chain reaction and template DNA derived from nullisomic strains to directly assign micronuclear DNA sequences to specific micronuclear chromosomes. Using this technique, a number of unique sequences and short repetitive sequences flanked by unique sequences have been mapped to four of the five germinal chromosomes.

scholarly journals Panhandle Polymerase Chain Reaction Amplifies MLL Genomic Translocation Breakpoint Involving Unknown Partner Gene

Blood ◽  
1997 ◽  
Vol 90 (12) ◽  
pp. 4679-4686 ◽  
Author(s):  
Carolyn A. Felix ◽  
Caroline S. Kim ◽  
Maureen D. Megonigal ◽  
Diana J. Slater ◽  
Douglas H. Jones ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lymphoblastic Leukemia ◽  
Repetitive Sequences ◽  
Translocation Breakpoint ◽  
Chromosome Band ◽  
Chain Reaction ◽  
Metaphase Chromosomes ◽  
Partner Gene ◽  
Translocation Breakpoints ◽  
Polymerase Chain

Abstract We used a new approach called panhandle polymerase chain reaction (PCR) to clone an MLL genomic translocation breakpoint in a case of acute lymphoblastic leukemia of infancy in which karyotype analysis was technically unsuccessful and did not show the translocation partner. Panhandle PCR amplified known MLL sequence 5′ of the breakpoint and 3′ sequence from the unknown partner gene from a DNA template with an intrastrand loop schematically shaped like a pan with a handle. The 7-kb panhandle PCR product contained the translocation breakpoint in MLL intron 8. The partner DNA included unique nonrepetitive sequences, Alu and mammalian apparent LTR-retrotransposon (MaLR) repetitive sequences, and a region of homology to expressed sequence tags. MaLR sequences have not been found before near leukemia-associated translocation breakpoints. The nonrepetitive sequences were not homologous to known partner genes of MLL. Screening of somatic cell hybrid and radiation hybrid lines by PCR and fluorescence in situ hybridization analysis of normal metaphase chromosomes mapped the partner DNA to chromosome band 4q21. Reverse transcriptase-PCR identified an MLL-AF-4 chimeric mRNA, indicating that panhandle PCR identified a fusion of MLL with a previously uncharacterized AF-4 intronic sequence. Panhandle PCR facilitates cloning translocation breakpoints and identifying unknown partner genes.

scholarly journals Single step polymerase chain reaction (PCR) for the diagnosis of the Leishmania (Viannia) subgenus

2004 ◽  
Vol 46 (6) ◽  
pp. 335-338 ◽  
Author(s):  
Byanca Regina Paiva ◽  
Luciana Neves Passos ◽  
Aloisio Falqueto ◽  
Rosely dos S. Malafronte ◽  
Heitor Franco de Andrade Jr.
Keyword(s):  
Polymerase Chain Reaction ◽  
Diagnostic Tool ◽  
Repetitive Sequences ◽  
Etiologic Agent ◽  
Single Step ◽  
Chain Reaction ◽  
Parasitological Diagnosis ◽  
Mucosal Involvement ◽  
Polymerase Chain ◽  
Step Polymerase Chain Reaction

In Brazil, the main etiologic agent of Leishmaniasis that frequently presents with mucosal involvement belongs to the Viannia subgenus. The therapeutic conduct in this disease depends on the parasitological diagnosis, and classical methods are restricted in identifying the agent. In this paper we describe a polymerase chain reaction (PCR), which uses primers designed from mini-exons repetitive sequences. The PCR amplifies a 177bp fragment that can distinguish (Viannia) from (Leishmania) subgenus. This test could be a useful diagnostic tool.

Sex determination of porcine embryos using a new developed duplex polymerase chain reaction procedure based on the amplification of repetitive sequences

2013 ◽  
Vol 25 (2) ◽  
pp. 417 ◽  
Author(s):  
Eva Torner ◽  
Eva Bussalleu ◽  
M. Dolors Briz ◽  
Alfonso Gutiérrez-Adán ◽  
Sergi Bonet
Keyword(s):  
Polymerase Chain Reaction ◽  
Single Cells ◽  
Repetitive Sequences ◽  
Repeat Sequence ◽  
Rrna Gene ◽  
Duplex Pcr ◽  
Chain Reaction ◽  
Cell Stage ◽  
Polymerase Chain ◽  
Primer Sets

Polymerase chain reaction (PCR)-based assays have become increasingly prevalent for sexing embryos. The aim of the present study was to develop a suitable duplex PCR procedure based on the amplification of porcine repetitive sequences for sexing porcine tissues, embryos and single cells. Primers were designed targeting the X12696 Y chromosome-specific repeat sequence (SUSYa and SUSYb; sex-related primer sets), the multicopy porcine-specific mitochondrial 12S rRNA gene (SUS12S; control primer set) and the X51555 1 chromosome repeat sequence (SUS1; control primer set). The specificity of the primer sets was established and the technique was optimised by testing combinations of two specific primer sets (SUSYa/SUS12S; SUSYb/SUS12S), different primer concentrations, two sources of DNA polymerase, different melting temperatures and different numbers of amplification cycles using genomic DNA from porcine ovarian and testicular tissue. The optimised SUSYa/SUS12S- and SUSYb/SUS12S-based duplex PCR procedures were applied to porcine in vitro-produced (IVP) blastocysts, cell-stage embryos and oocytes. The SUSYb/SUS12S primer-based procedure successfully sexed porcine single cells and IVP cell-stage embryos (100% efficiency), as well as blastocysts (96.6% accuracy; 96.7% efficiency). This is the first report to demonstrate the applicability of these repetitive sequences for this purpose. In conclusion, the SUSYb/SUS12S primer-based duplex PCR procedure is highly reliable and sensitive for sexing porcine IVP embryos.

scholarly journals Panhandle Polymerase Chain Reaction Amplifies MLL Genomic Translocation Breakpoint Involving Unknown Partner Gene

Blood ◽  
1997 ◽  
Vol 90 (12) ◽  
pp. 4679-4686 ◽  
Author(s):  
Carolyn A. Felix ◽  
Caroline S. Kim ◽  
Maureen D. Megonigal ◽  
Diana J. Slater ◽  
Douglas H. Jones ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Lymphoblastic Leukemia ◽  
Repetitive Sequences ◽  
Translocation Breakpoint ◽  
Chromosome Band ◽  
Chain Reaction ◽  
Metaphase Chromosomes ◽  
Partner Gene ◽  
Translocation Breakpoints ◽  
Polymerase Chain

We used a new approach called panhandle polymerase chain reaction (PCR) to clone an MLL genomic translocation breakpoint in a case of acute lymphoblastic leukemia of infancy in which karyotype analysis was technically unsuccessful and did not show the translocation partner. Panhandle PCR amplified known MLL sequence 5′ of the breakpoint and 3′ sequence from the unknown partner gene from a DNA template with an intrastrand loop schematically shaped like a pan with a handle. The 7-kb panhandle PCR product contained the translocation breakpoint in MLL intron 8. The partner DNA included unique nonrepetitive sequences, Alu and mammalian apparent LTR-retrotransposon (MaLR) repetitive sequences, and a region of homology to expressed sequence tags. MaLR sequences have not been found before near leukemia-associated translocation breakpoints. The nonrepetitive sequences were not homologous to known partner genes of MLL. Screening of somatic cell hybrid and radiation hybrid lines by PCR and fluorescence in situ hybridization analysis of normal metaphase chromosomes mapped the partner DNA to chromosome band 4q21. Reverse transcriptase-PCR identified an MLL-AF-4 chimeric mRNA, indicating that panhandle PCR identified a fusion of MLL with a previously uncharacterized AF-4 intronic sequence. Panhandle PCR facilitates cloning translocation breakpoints and identifying unknown partner genes.

Detection of repetitive sequences ofTrichinella spiralis by the polymerase chain reaction in experimentally infected mice

1991 ◽  
Vol 77 (2) ◽  
pp. 180-182 ◽  
Author(s):  
Jean Dupouy-Camet ◽  
Claude Soulé ◽  
Jean-Pierre Guillou ◽  
Evelyne Rouer ◽  
Sandra Lavareda de Souza ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Repetitive Sequences ◽  
Chain Reaction ◽  
Polymerase Chain
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