A two‐buffer method used for both polymerase chain reaction product purification and DNA gel extraction

2019 ◽  
Vol 94 (9) ◽  
pp. 2928-2934 ◽  
Author(s):  
Zihui Wu ◽  
Houliang Zhao ◽  
Cheng Zhang ◽  
Juan Wang ◽  
Shasha Zhao ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Product ◽  
Chain Reaction ◽  
Polymerase Chain

Detection and identification of Clavibacter michiganensis subsp. sepedonicus and Clavibacter michiganensis subsp. michiganensis by nonradioactive hybridization, polymerase chain reaction, and restriction enzyme analysis

1994 ◽  
Vol 40 (12) ◽  
pp. 1007-1018 ◽  
Author(s):  
J. L. W. Rademaker ◽  
J. D. Janse
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Product ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Bacterial Strains ◽  
Clavibacter Michiganensis ◽  
Chain Reaction ◽  
Clavibacter Michiganensis Subsp ◽  
Detection And Identification ◽  
Polymerase Chain

To develop a rapid and reliable detection and identification method for Clavibacter michiganensis subsp. sepedonicus and C. michiganensis subsp. michiganensis, two biotinylated probes and derived primer sets were evaluated for specificity using a large number of bacterial strains. Detection in dot blot analysis using the Diagen probe against C. michiganensis subsp. sepedonicus was possible with all 32 C. michiganensis subsp. sepedonicus strains tested. Cross-hybridization occurred with all nine C. michiganensis subsp. insidiosus strains tested. No hybridization occurred with any of 54 other related and unrelated bacterial strains including C. michiganensis subsp. michiganensis, C. michiganensis subsp. nebraskensis, C. michiganensis subsp. tessellarius, C. iranicus, C. rathayi, and C. tritici and potato saprophytes. Hybridization of the MIC 1 probe against C. michiganensis subsp. michiganensis was obtained with 22 out of 24 C. michiganensis subsp. michiganensis strains. A weak hybridization signal occurred only with two strains of C. michiganensis subsp. insidiosns. No hybridization occurred with any of the 71 other related and unrelated bacterial strains tested including tomato saprophytes. Restriction fragment length polymorphisms detected with the Diagen probe allowed differentiation between C. michiganensis subsp. sepedonicus and the related C. michiganensis subsp. insidiosus. Restriction fragment length polymorphism analysis using the MIC 1 probe and BamH1 showed at least two groups of patterns within C. michiganensis subsp. michiganensis. By using a primer set derived from the Diagen probe, a DNA sequence could be amplified with all C. michiganensis subsp. sepedonicus strains tested. Only the nontarget organism C. michiganensis subsp. insidiosus yielded a similar polymerase chain reaction product. Restriction enzyme analysis of the polymerase chain reaction product enabled rapid distinction between the subspecies. With a CMM primer set derived from the MIC 1 probe a DNA sequence was amplified from the same 22 out of 24 C. michiganensis subsp. michiganensis strains that showed hybridization with the MIC 1 probe. The polymerase chain reaction product could be verified by restriction enzyme analysis. The Diagen and MIC 1 probes and derived primer sets were shown to be useful for the detection and identification of C. michiganensis subsp. sepedonicus and C. michiganensis subsp. michiganensis. The MIC 1 probe, however, failed to detect two strains of the latter subspecies.Key words: biotin, PCR, REA, potato bacterial ring rot, bacterial canker of tomato, RFLP, Clavibacter michiganensis subsp. insidiosus.

Real-Time Homogeneous Assay of Rapid Cycle Polymerase Chain Reaction Product for Identification ofLeptonema illini

1998 ◽  
Vol 259 (1) ◽  
pp. 112-117 ◽  
Author(s):  
Tony H.S. Woo ◽  
Bharat K.C. Patel ◽  
Marina Cinco ◽  
Lee D. Smythe ◽  
Meegan L. Symonds ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Polymerase Chain Reaction Product ◽  
Chain Reaction ◽  
Homogeneous Assay ◽  
Polymerase Chain

Automated polymerase chain reaction product sample preparation for capillary electrophoresis analysis

1996 ◽  
Vol 683 (1) ◽  
pp. 109-114 ◽  
Author(s):  
Phillip Belgrader ◽  
Joseph M. Devaney ◽  
Susie A. Del Rio ◽  
Kelly A. Turner ◽  
Kristal R. Weaver ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Capillary Electrophoresis ◽  
Sample Preparation ◽  
Polymerase Chain Reaction Product ◽  
Chain Reaction ◽  
Polymerase Chain

scholarly journals MORPHOLOGICAL AND MOLECULAR IDENTIFICATION OF ZOONOTIC TREMATODE METACERCARIAE (Haplorchis taitui) IN FRESHWATER FISH IN IRAN

2021 ◽  
Vol 28 ◽  
pp. 1-9
Author(s):  
Alireza Golchin Manshadi
Keyword(s):  
Public Health ◽  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Product ◽  
Deoxyribonucleic Acid ◽  
Nucleotide Sequence Analysis ◽  
Chain Reaction ◽  
Mean Intensity ◽  
The Mean ◽  
Polymerase Chain ◽  
Haplorchis Taichui

The intestinal trematode, Haplorchis taichui (Nishigori, 1924), is an important in public health that causes infection in humans and animals especially in Asia and in parts of Africa and the Americas. Haplorchis taitui metacercariae were found in the gills of Cyprinion macrostomus (Heckel) and Capoeta barroisi persica (Karaman) collected from the Shapour River. Morphological excysted metacercariae were identified as wet mounts under a stereomicroscope. Then, the samples were subjected to molecular analysis. The result showed that 69% of examined fish (n = 30) were diagnosed infected with encysted metacercariae in gills. The mean intensity was 8.3 ± 16.9 parasites per fish. The morphometrical values agree with the findings of other studies with the small differences and polymerase chain reaction product length and nucleotide sequence analysis of 18S ribosomal deoxyribonucleic acid gene showed a similarity of over 99% between the specimens and the Haplorchis taichui (Nishigori, 1924) recorded in GenBank.

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