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

Plants ◽  
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.

scholarly journals Species Identification of Crassostrea and Ostrea Oysters by Polymerase Chain Reaction Amplification of the 5S rRNA Gene

2006 ◽  
Vol 89 (1) ◽  
pp. 144-148 ◽  
Author(s):  
Ismael Cross ◽  
Laureana Rebordinos ◽  
Edgardo Diaz
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Amplification ◽  
5S Rdna ◽  
Universal Primers ◽  
Rrna Gene ◽  
Chain Reaction ◽  
Nontranscribed Spacer ◽  
Crassostrea Angulata ◽  
Polymerase Chain ◽  
Species Specific

Abstract A specific multiplex polymerase chain reaction (PCR) was developed for the identification of Crassostrea angulata, C. gigas, Ostrea edulis, and O. stentina oyster species. Universal primers were used for the amplification of complete repetition units of 5S rDNA in each of the 4 species. The alignment of the obtained sequences was the basis for the specific design of species-specific primers (ED1, ED2, ST1, ST2, CR1, and CR2) located in the nontranscribed spacer regions. The different sizes of the species-specific amplicons, separated by agarose gel electrophoresis, allowed identification of Crassostrea and Ostrea species. A multiplex PCR with a set of the 6 designed primers showed that they did not interfere with each other and bound specifically to the DNA target. This genetic marker can be very useful for traceability of the species, application in the management of oyster cultures, and conservation of the genetic resources of the species.

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

Genome ◽  
2012 ◽  
Vol 55 (5) ◽  
pp. 370-382 ◽  
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.

Routine identification of four sympatric species of calanoid copepods Pseudocalanus spp. in the Atlantic Arctic using a species-specific polymerase chain reaction

2020 ◽  
Vol 48 (1) ◽  
pp. 62-72
Author(s):  
E. A. Ershova
Keyword(s):  
Polymerase Chain Reaction ◽  
Life Cycles ◽  
The Arctic ◽  
Relative Distribution ◽  
Chain Reaction ◽  
Specific Polymerase Chain Reaction ◽  
Routine Identification ◽  
Polymerase Chain ◽  
Species Specific ◽  
Plankton Communities

Сalanoid copepods of the genus Pseudocalanus play an important role in the plankton communities of the Arctic and boreal seas, often dominating in numbers and constituting a significant proportion of the biomass of zooplankton. Despite their high presence and significance in the shelf plankton communities, species-specific studies of the biology of these are significantly hampered by extremely small morphological differences between them, especially at the juvenile stages, at which they are virtually indistinguishable. In this paper, we describe a new, routine and low-cost molecular method for identifying all Pseudocalanus species found in the Atlantic sector of the Arctic: the Arctic P. acuspes, P. minutus and the boreal P. moultoni and P. elongatus, and apply it to describe the relative distribution of these species in four locations of the Arctic and sub-Arctic. With this method, species-specific polymerase chain reaction (ssPCR), mass identification of individuals of any developmental stage, including nauplii, is possible. This method can serve as an excellent tool for studying the species-specific biology of this group, describing their life cycles, as well as monitoring changes in Arctic marine ecosystems under the influence of changing climate.

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

1993 ◽  
Vol 39 (9) ◽  
pp. 1927-1933 ◽  
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.

Polymerase Chain Reaction Diagnosis of Leishmaniasis: A Species-Specific Approach

2016 ◽  
pp. 113-124 ◽  
Author(s):  
Eglys González-Marcano ◽  
Hirotomo Kato ◽  
Juan Luis Concepción ◽  
María Elizabeth Márquez ◽  
Alberto Paniz Mondolfi
Keyword(s):  
Polymerase Chain Reaction ◽  
Chain Reaction ◽  
Specific Approach ◽  
Polymerase Chain ◽  
Species Specific
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