scholarly journals Monovalent Copper and Silver Ions Block DNA Polymerase Chain Reaction

Author(s):  
Lea Moshkovich ◽  
Oshra Saphier ◽  
Stanislav Popov ◽  
Yoram Shotland ◽  
Eldad Silberstein ◽  
...  

Abstract In the present study we present the dramatic effect that monovalent copper ions (Cu(I))have on the DNA polymerase chain reaction, and the moderate effect which monovalent silver ions (Ag(I)) have on it. Our research utilizes the commercial Polymerase Chain Reaction (PCR) system: in anaerobic conditions, in the presence of less than 0.1 mM of Cu(I) ions or in the presence of less than 10mM of Ag(I) ions, the PCR system was entirely shut down.Under the same conditions, 1 mM of divalent copper ions (Cu(II)) ions shows only a minor effect, while10mM of divalent Ni and Zn ions shows no effect at all.This finding can give some explanation for the strong antimicrobial activity of monovalent copper ions (Cu(I))as well as Ag(I). Although the mechanism of this effect is not yet fully understood, we recently published results showing that under the conditions of acidic pH, an unfavorable carbon source, low molecular oxygen concentration and elevated temperatures, the antibacterial action of Cu(I) ions is boosted, with a 106 bacterial population eliminated in less than 1 min by 0.4mM of Cu(I). Microscopy checking of E.coli morphology and light scattering testes showed mortality of bacteria with almost no lysis. These results suggest that rapid and lethal metabolic damage is the main mechanism of Cu(I)’s antimicrobial effect.

Agrikultura ◽  
2010 ◽  
Vol 21 (1) ◽  
Author(s):  
Nono Carsono ◽  
Sri Nurlianti ◽  
Inez Nur Indrayani ◽  
Ade Ismail ◽  
Tri Joko Santoso ◽  
...  

Transformasi gen Glu-1Dx5, pengendali utama karakter elastisitas dan daya mengembang adonan dari gandum, telah berhasil ditransfer ke dalam genom tanaman padi kultivar Fatmawati dengan menggunakan penembakan partikel, dengan tujuan untuk memperbaiki kualitas adonan tepung beras. Galur-galur harapan telah diperoleh, tetapi karena telah mengalami penyerbukan sendiri selama 1-2 generasi yang menyebabkan transgen mengalami segregasi, maka diperlukan upaya pendeteksian transgen pada populasi putative transgenik ini. Upaya ini dapat dilakukan, antara lain dengan menggunakan teknik Polymerase Chain Reaction (PCR) yang memungkinkan perbanyakan fragmen DNA yang spesifik (gen) secara cepat dalam jumlah banyak.  Percobaan ini bertujuan untuk mendapatkan tanaman padi transgenik yang memiliki gen Glu-1Dx5 pada dua generasi yang sedang bersegregasi. DNA genom dari 149 tanaman padi (generasi T1 sebanyak 14 tanaman, generasi T2 sebanyak 134 tanaman, dan satu tanaman non-transgenik) telah diekstraksi menggunakan Genomic DNA Purification Kit dari Fermentas. Plasmid pK+Dx5 digunakan sebagai positif kontrol, selain itu digunakan juga enzim Taq DNA polymerase dari Go Green Taq® Master Mix (Promega) dan 2 primer spesifik yang mengamplifikasi coding region dari Glu-1Dx5 (2,5 kb). Hasil percobaan menunjukkan, tanaman padi yang memiliki gen Glu-1Dx5 pada generasi T2-7 sebanyak 26 tanaman, T2-11 : 12 tanaman, T2-12 : 3 tanaman, T2-40 : 3 tanaman dan T2-45 : 5 tanaman. Seluruh tanaman generasi T1 tidak memiliki insert. Hasil ini menunjukkan bahwa gen Glu-1Dx5 sudah terintegrasi ke dalam genom tanaman padi kultivar Fatmawati dan diwariskan dari satu generasi ke generasi berikutnya.


RSC Advances ◽  
2013 ◽  
Vol 3 (43) ◽  
pp. 20793 ◽  
Author(s):  
Samir Mandal ◽  
Maidul Hossain ◽  
T. Muruganandan ◽  
Gopinatha Suresh Kumar ◽  
Keya Chaudhuri

2016 ◽  
Author(s):  
vikash bhardwaj ◽  
Vikash bhardwaj ◽  
Kulbhushan Sharma

Vox Sanguinis ◽  
1997 ◽  
Vol 72 (3) ◽  
pp. 192-196 ◽  
Author(s):  
Dong-Feng Chen ◽  
Ladislav T. Pastucha ◽  
Hai-Yen Chen ◽  
Janos G. Kadar ◽  
Walter Stangel

Genome ◽  
1993 ◽  
Vol 36 (6) ◽  
pp. 1029-1031 ◽  
Author(s):  
Juan Manuel González ◽  
Esther Ferrer

Random amplified polymorphic DNA analysis was performed by applying a set of 13 arbitrary 10-mer primers to 19 Hordeum species and subspecies. High levels of variation in fragment pattern were observed both within and among species with most of the primers used. Genetic similarities between accessions and species were calculated from the fragment patterns. The resulting phenograms confirmed previous relationships among the Hordeum species.Key words: random amplified polymorphic DNA, polymerase chain reaction, polymorphism, Hordeum.


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