"Portraying" of plant genomes using polymerase chain reaction amplification of ribosomal 5S genes

Nontranscribed spacers of plant genes coding for ribosomal 5S RNA were amplified using the polymerase chain reaction. Primers were synthesized that were complementary to 3′ (direct) and 5′ (reverse) ends of the coding region and that are universal for higher plants. The patterns of polymerase chain reaction products are species and, sometimes, variety specific. The use of this approach for identification of barley 5S genes in chromosome-addition lines of wheat is discussed. This principle can be applied for the "portraying" of other tandem repetitive genes containing divergent regions.Key words: polymerase chain reaction, 5S genes, plants, DNA polymorphism.

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Rye chromosome-specific polymerase chain reaction products developed by primers designed from the EcoO109I recognition site

Genome ◽

10.1139/g2012-024 ◽

2012 ◽

Vol 55 (5) ◽

pp. 370-382 ◽

Cited By ~ 7

Author(s):

Motonori Tomita ◽

Akiyoshi Seno

Keyword(s):

Polymerase Chain Reaction ◽

Dna Markers ◽

Addition Lines ◽

Reaction Products ◽

Chain Reaction ◽

Nylon Membrane ◽

Chromosome Addition ◽

Chromosome Addition Lines ◽

Pcr Products ◽

Polymerase Chain

From our analysis of repeat sequences in the rye genome, the presence of multiple restriction sites of EcoO109I (5′–PuGGNCCPy–3′) across the genome has been predicted. By first using primers designed to contain EcoO109I sites in polymerase chain reaction (PCR), polymorphic DNA markers were effectively obtained. A total of 43 types of 10-mer primers containing EcoO109I sites were applied for PCR by using genomic DNA of Secale cereale self-fertile line IR27 and Triticum aestivum ‘Chinese Spring’ (CS) as the template. Twenty two primers detected polymorphisms between wheat and rye, and they were applied for PCR using a series of CS wheat – ‘Imperial’ rye chromosome addition lines as templates. Nine chromosome-specific amplification fragments identified on five chromosomes were collected from gels and hybridized with nylon membrane-transferred PCR products from the wheat–rye chromosome addition lines. The gel blot was only observed between the collected fragments; therefore, these fragments were confirmed to be chromosome-specific. These fragments were sequenced and converted to sequence-tagged site (STS) primers. We therefore introduce a new method for building chromosome-specific DNA markers: (i) multiple polymorphic fragments can be obtained from EcoO109I primers and (ii) the addition of three nucleotides to the EcoO109I site restricts the amplification region to generate chromosome-specific fragments.

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Determination the causative strain for hydatid cyst in Iraqi cattle by using ND1 gene

The Iraqi Journal of Veterinary Medicine ◽

10.30539/iraqijvm.v41i1.71 ◽

2017 ◽

Vol 41 (1) ◽

pp. 11-16

Author(s):

Mohammed J. Muhaidi

Keyword(s):

Polymerase Chain Reaction ◽

Nadh Dehydrogenase ◽

Sequence Data ◽

Polymerase Chain Reaction Amplification ◽

Acid Extraction ◽

Reaction Products ◽

Hydatid Cysts ◽

Chain Reaction ◽

Polymerase Chain ◽

Sheep Strain

Hydatid Cysts were obtained from 15 cows from liver, lung, spleen, heart, and peritoneal cavity, between December 2014 and October 2015. Hydatid cysts (protoscoleces) were used for deoxyribonucleic acid extraction by using mechanical grinder. The purification of mtDNA was done by (promega kit, USA). The mitochondrial NADH dehydrogenase subunit 1 (ND1) genes was used as targets for polymerase chain reaction amplification, all hydatid cysts yielded amplification products. Polymerase chain reaction product for NADH1 800 basic pair. The polymerase chain reaction products were purified and partial sequences were generated. The sequences obtained were found to align with corresponding region for ND1 gene in the Gene Bank nucleotide database confirming to genotype of sheep strain (G1) in Iraq, Phylogenetic analysis of partial sequence data from ND1 genes for obtained Phylogenetic tree. G1 genotype was the most common taxon and the actual source of infection of Iraqi's cattle. All of 15 strains were G1 genotype (sheep strain) based on the partial sequences of NADH dehydrogenase 1 (ND1).

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Identification of a single base change in a new human mutant glucose-6-phosphate dehydrogenase gene by polymerase-chain-reaction amplification of the entire coding region from genomic DNA

Biochemical Journal ◽

10.1042/bj2710157 ◽

1990 ◽

Vol 271 (1) ◽

pp. 157-160 ◽

Cited By ~ 49

Author(s):

V Poggi ◽

M Town ◽

N S Foulkes ◽

L Luzzatto

Keyword(s):

Polymerase Chain Reaction ◽

Genomic Dna ◽

Polymerase Chain Reaction Amplification ◽

Base Change ◽

Nucleotide Position ◽

Coding Region ◽

Chain Reaction ◽

G6pd Gene ◽

Polymerase Chain ◽

Glucose 6 Phosphate Dehydrogenase

We report the characterization at the molecular level of a mutant glucose-6-phosphate dehydrogenase (G6PD) gene in a Greek boy who presented with a chronic non-spherocytic haemolytic anaemia. In order to identify the mutation from a small amount of patient material, we adopted an approach which by-passes the need to construct a library by using the polymerase chain reaction. The entire coding region was amplified in eight sections, with genomic DNA as template. The DNA fragments were then cloned in an M13 vector and sequenced. The only difference from the sequence of normal G6PD was a T----G substitution at nucleotide position 648 in exon 7, which predicts a substitution of leucine for phenylalanine at amino acid position 216. This mutation creates a new recognition site for the restriction nuclease BalI. We confirmed the presence of the mutation in the DNA of the patient's mother, who was found to be heterozygous for the new BalI site. This is the first transversion among the point mutations thus far reported in the human G6PD gene.

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Polymerase Chain Reaction Amplification of Archival Material for Parvovirus B19 in Children with Transient Erythroblastopenia of Childhood

Fetal and Pediatric Pathology ◽

10.3109/15513819609168684 ◽

1996 ◽

Vol 16 (3) ◽

pp. 471-478 ◽

Cited By ~ 8

Author(s):

Beverly Barton Rogers ◽

Zora Rogers ◽

Charles Timmons

Keyword(s):

Polymerase Chain Reaction ◽

Polymerase Chain Reaction Amplification ◽

Parvovirus B19 ◽

Chain Reaction ◽

Archival Material ◽

Reaction Amplification ◽

Polymerase Chain

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Faculty Opinions recommendation of Synthesis and polymerase chain reaction amplification of DNA strands containing an unnatural triazole linkage.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1158100.618201 ◽

2009 ◽

Author(s):

Scott Silverman

Keyword(s):

Polymerase Chain Reaction ◽

Polymerase Chain Reaction Amplification ◽

Chain Reaction ◽

Dna Strands ◽

Reaction Amplification ◽

Polymerase Chain

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Polymerase chain reaction amplification and typing of rotavirus in environmental samples

Water Science & Technology ◽

10.2166/wst.1995.0643 ◽

1995 ◽

Vol 31 (5-6) ◽

pp. 371-374 ◽

Cited By ~ 5

Author(s):

R. Gajardo ◽

R. M. Pintó ◽

A. Bosch

Keyword(s):

Polymerase Chain Reaction ◽

Environmental Samples ◽

Polymerase Chain Reaction Amplification ◽

Rt Pcr ◽

Chain Reaction ◽

Pcr Assay ◽

Group A ◽

Reaction Amplification ◽

Stool Specimens ◽

Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

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