A PCR assay for detection of a 2RL.2BS wheat–rye chromosome translocation

A 2RL.2BS wheat–rye translocation, present in the wheat germplasm line Hamlet, carries a gene for resistance to Hessian fly biotype L, one of the most virulent biotypes presently encountered in wheat production environments. Unlike several other wheat–rye chromosome translocations common in wheat breeding programs, 2RL lacks genes encoding storage proteins or other easily selected markers. Oligonucleotide primers synthesized from published sequences derived from the R173 family of moderately repetitive rye DNA were used in the DNA polymerase chain reaction (PCR) to identify specific markers for 2RL. The same primers, when used with DNA extracted from additional wheat–rye translocation lines of importance to the wheat breeding community, gave distinctive PCR products for each genotype. The single primer pair, PAWS5 and PAWS6, may, therefore, have wide applicability for the identification of wheat–rye chromosomal translocations presently encountered in wheat breeding populations. Key words : 2RL.2BS wheat–rye chromosome translocation, polymerase chain reaction, detection.

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Variability among members of the Hor-2 multigene family

Genome ◽

10.1139/g93-055 ◽

1993 ◽

Vol 36 (3) ◽

pp. 397-403 ◽

Cited By ~ 17

Author(s):

Vladimir Kanazin ◽

Evgeny Ananiev ◽

Tom Blake

Keyword(s):

Polymerase Chain Reaction ◽

Gene Family ◽

Storage Proteins ◽

Gene Families ◽

Reaction Sequence ◽

Chain Reaction ◽

Genes Encoding ◽

Polymerase Chain ◽

Encoding Gene ◽

Barley Genotypes

The hordeins comprise the major prolamin storage proteins of barley. Two major and one minor gene families encode these alcohol-soluble proteins. The Hor-2 gene family encoding the B-hordeins has been estimated to contain 15–30 copies. Although several genes encoding B-hordeins have been cloned and sequenced, little is known about the mechanisms responsible for the generation of the enormous genetic variability at this locus. Polymerase chain reaction sequence amplification provided a simple technique that permitted the amplification of the Hor-2 gene family members from the genomes of several barley genotypes. Sequence analysis of clones permitted the identification of a region within the Hor-2 structural gene that appears to undergo recombinational and slippage-like gene conversion events. In this report we describe variability of the B-hordein genes, possible mechanisms responsible for it, and implications this may have on the evolution of prolamin-encoding gene families.Key words: barley, hordeins, polymerase chain reaction, polymorphism.

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A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽

10.3390/plants10010004 ◽

2020 ◽

Vol 10 (1) ◽

pp. 4

Author(s):

Oleg S. Alexandrov ◽

Olga V. Razumova ◽

Gennady I. Karlov

Keyword(s):

Polymerase Chain Reaction ◽

Molecular Markers ◽

Dna Markers ◽

5S Rdna ◽

Chain Reaction ◽

Closely Related Species ◽

Pcr Products ◽

Polymerase Chain ◽

Species Specific ◽

Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

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Automated closed-vessel system for in vitro diagnostics based on polymerase chain reaction

Clinical Chemistry ◽

10.1093/clinchem/39.9.1927 ◽

1993 ◽

Vol 39 (9) ◽

pp. 1927-1933 ◽

Cited By ~ 39

Author(s):

J B Findlay ◽

S M Atwood ◽

L Bergmeyer ◽

J Chemelli ◽

K Christy ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Pcr Amplification ◽

Automated System ◽

Closed Vessel ◽

Chain Reaction ◽

Panel Testing ◽

Pcr Product ◽

Pcr Products ◽

Polymerase Chain

Abstract An automated system for polymerase chain reaction (PCR) amplification and detection combats false-positive results caused by "PCR product carryover." The system uses a single vessel for both PCR amplification and the subsequent detection of PCR products, eliminating the need to handle PCR products in an open environment and risk product carryover. The sample and PCR reagents are introduced into one compartment within the vessel, and amplification occurs as they are thermally cycled. Other compartments contain the reagents for detection of PCR products. Pressure from a roller provides for sequential delivery of the contents of the compartments to a detection area. The PCR products are biotinylated at their 5' ends during amplification through the use of biotinylated primers. After delivery to the detection area, they are specifically captured by hybridization with immobilized oligonucleotide probes. Subsequent reaction with streptavidin-horseradish peroxidase conjugate forms a complex that catalyzes dye formation from dye precursor. Wash steps minimize nonspecific background. This format is amenable to multiplexing, permitting internal controls, speciation of bacteria, typing of viruses, and panel testing. An HIV assay performed with this system demonstrated 100% sensitivity and 95% specificity for 64 patients' samples relative to a conventional PCR assay based on 32P solution hybridization. Similarly, an automated closed-vessel assay of cytomegalovirus exhibited 97.5% sensitivity and 100% specificity.

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Fluorometric Polymerase Chain Reaction (PCR) Enzyme-Linked Immunosorbent Assay for Quantification of Immuno-PCR Products in Microplates

Analytical Biochemistry ◽

10.1006/abio.1996.9989 ◽

1997 ◽

Vol 246 (1) ◽

pp. 140-145 ◽

Cited By ~ 61

Author(s):

Christof M. Niemeyer ◽

Michael Adler ◽

Dietmar Blohm

Keyword(s):

Polymerase Chain Reaction ◽

Immunosorbent Assay ◽

Enzyme Linked Immunosorbent Assay ◽

Chain Reaction ◽

Pcr Products ◽

Polymerase Chain

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PCR-induced (ligase-free) subcloning: a rapid reliable method to subclone polymerase chain reaction (PCR) products

Nucleic Acids Research ◽

10.1093/nar/18.7.1920 ◽

1990 ◽

Vol 18 (7) ◽

pp. 1920-1920 ◽

Cited By ~ 36

Author(s):

Alan R. Shuldiner ◽

Laurie A. Scott ◽

Jesse Roth

Keyword(s):

Polymerase Chain Reaction ◽

Reliable Method ◽

Chain Reaction ◽

Pcr Products ◽

Polymerase Chain

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Detection of the chromosomal translocation t(11;14) by polymerase chain reaction in mantle cell lymphomas

Blood ◽

10.1182/blood.v83.7.1871.1871 ◽

1994 ◽

Vol 83 (7) ◽

pp. 1871-1875 ◽

Cited By ~ 150

Author(s):

R Rimokh ◽

F Berger ◽

G Delsol ◽

I Digonnet ◽

JP Rouault ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Chromosomal Translocation ◽

Nucleotide Sequencing ◽

Chromosome 11 ◽

Chromosome 14 ◽

Chain Reaction ◽

Mantle Cell ◽

Pcr Products ◽

Polymerase Chain ◽

Ig Heavy Chain

Abstract The t(11;14)(q13;q32) and its molecular counterpart, BCL1 rearrangement, are consistent features of mantle cell lymphoma (MCL). Rearrangement is thought to deregulate the nearby CCND1 (BCL1/PRAD1) proto-oncogene, a member of the cyclin G1 gene family, and thereby to contribute to tumorigenesis. We and others have previously shown that the BCL1 locus is rearranged in 55% to 60% of MCL patients and that, on chromosome 11, more than 80% of the breakpoints are localized within a 1-kbp DNA segment known as the major translocation cluster (MTC). We have determined the nucleotide sequence for a portion of the MTC region, and constructed chromosome 11-specific oligonucleotides that were in conjunction with a consensus immunoglobulin (Ig) heavy chain joining region (JH) primer used to perform the polymerase chain reaction (PCR) to amplify t(11;14) chromosomal junctional sequences in DNA from 16 MCL patients with breakpoints in the MTC region. 15 of the 16 breakpoints that occurred at the MTC region were amenable to PCR detection. The sizes of the amplified bands, the existence or not of a Sac I site in the PCR products, and nucleotide sequencing of the amplified DNA from four patients showed that the breakpoints share a remarkable tendency to tightly cluster within 300 bp on chromosome 11, some of them occurring at the same nucleotide. On chromosome 14, the breakpoints were localized within the Ig JH. Our findings indicate that a BCL1 rearrangement can be detected using this approach in roughly one half of the MCL patients. This has implications for both the diagnosis and the clinical management of MCL.

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Detection of Subclinical Systemic Disease in Primary CNS Lymphoma by Polymerase Chain Reaction of the Rearranged Immunoglobulin Heavy-Chain Genes

Journal of Clinical Oncology ◽

10.1200/jco.2006.06.7165 ◽

2006 ◽

Vol 24 (29) ◽

pp. 4754-4757 ◽

Cited By ~ 55

Author(s):

Kristoph Jahnke ◽

Michael Hummel ◽

Agnieszka Korfel ◽

Thomas Burmeister ◽

Philipp Kiewe ◽

...

Keyword(s):

Bone Marrow ◽

Polymerase Chain Reaction ◽

Systemic Disease ◽

Primary Cns Lymphoma ◽

Cns Lymphoma ◽

Chain Reaction ◽

Blood Samples ◽

Pcr Products ◽

Polymerase Chain

Purpose To search for subclinical systemic disease in bone marrow and peripheral blood in patients with primary CNS lymphoma (PCNSL) to elucidate whether extracerebral relapse may represent a sequel of initial occult systemic disease rather than true extracerebral spread. Patients and Methods Bone marrow and peripheral-blood specimens of 24 PCNSL patients were examined using polymerase chain reaction (PCR) for analysis of clonally rearranged immunoglobulin heavy-chain (IgH) genes. Results Identical dominant PCR products were found in bone marrow aspirates, blood samples, and tumor biopsy specimens of two patients, indicating that the same tumor cell population is present in the CNS and in extracerebral sites. Follow-up IgH PCR performed in one of these patients in complete remission 24 months after diagnosis yielded a persistent monoclonal product in the blood. An oligoclonal IgH rearrangement pattern was found in the tumor specimen of two other patients, whereas bone marrow and blood samples demonstrated the same dominant PCR products. Follow-up PCR showed a persistent monoclonal amplificate in blood in one of these patients 27 months after diagnosis. Conclusion It could be demonstrated for the first time that subclinical systemic disease can be present in PCNSL patients at initial diagnosis. Our findings may have an impact on the understanding of PCNSL pathogenesis and the extent of staging and treatment.

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