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Plos One : Sliding Over the Blocks in Enzyme-free Rna Copying – One-pot Primer Extension in Ice, Volume 8

By Barash, Danny

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Book Id: WPLBN0003966785
Format Type: PDF eBook :
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Reproduction Date: 2015

Title: Plos One : Sliding Over the Blocks in Enzyme-free Rna Copying – One-pot Primer Extension in Ice, Volume 8  
Author: Barash, Danny
Volume: Volume 8
Language: English
Subject: Journals, Science, Medical Science
Collections: Periodicals: Journal and Magazine Collection (Contemporary)
Historic
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Publisher: Plos

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Barash, D. (n.d.). Plos One : Sliding Over the Blocks in Enzyme-free Rna Copying – One-pot Primer Extension in Ice, Volume 8. Retrieved from http://ebook.worldlibrary.net/


Description
Description : Template-directed polymerization of RNA in the absence of enzymes is the basis for an information transfer in the ‘RNA-world’ hypothesis and in novel nucleic acid based technology. Previous investigations established that only cytidine rich strands are efficient templates in bulk aqueous solutions while a few specific sequences completely block the extension of hybridized primers. We show that a eutectic water/ice system can support Pb2+/Mg2+-ion catalyzed extension of a primer across such sequences, i.e. AA, AU and AG, in a one-pot synthesis. Using mixtures of imidazole activated nucleotide 5′-monophosphates, the two first “blocking” residues could be passed during template-directed polymerization, i.e., formation of triply extended products containing a high fraction of faithful copies was demonstrated. Across the AG sequence, a mismatch sequence was formed in similar amounts to the correct product due to U·G wobble pairing. Thus, the template-directed extension occurs both across pyrimidine and purine rich sequences and insertions of pyrimidines did not inhibit the subsequent insertions. Products were mainly formed with 2′-5′-phosphodiester linkages, however, the abundance of 3′–5′-linkages was higher than previously reported for pyrimidine insertions. When enzyme-free, template-directed RNA polymerization is performed in a eutectic water ice environment, various intrinsic reaction limitations observed in bulk solution can then be overcome.


 

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