scholarly journals Generation of long, fully modified, and serum-resistant oligonucleotides by rolling circle amplification

2015 ◽  
Vol 13 (38) ◽  
pp. 9820-9824 ◽  
Author(s):  
Marcel Hollenstein
Keyword(s):  
Rolling Circle Amplification ◽  
Single Stranded Dna ◽  
Rolling Circle ◽  
Dna Oligonucleotides ◽  
Amplification Method ◽  
Nucleoside Triphosphates

Nucleoside triphosphates modified at any level of the scaffold were shown to be compatible with the rolling circle amplification method. The combination of modified dNTPs and RCA enables the generation of long, fully modified, single-stranded DNA oligonucleotides.

scholarly journals An improved experimental pipeline for preparing circular ssDNA viruses for next-generation sequencing

2020 ◽  
Author(s):  
Catherine D. Aimone ◽  
J. Steen Hoyer ◽  
Anna E. Dye ◽  
David O. Deppong ◽  
Siobain Duffy ◽  
...  
Keyword(s):  
Next Generation Sequencing ◽  
Rolling Circle Amplification ◽  
Next Generation ◽  
Viral Dna ◽  
Single Stranded Dna ◽  
Dna Viruses ◽  
Short Read ◽  
Rolling Circle ◽  
Optimized Protocol ◽  
Generation Sequencing

AbstractWe present an optimized protocol for enhanced amplification and enrichment of viral DNA for Next Generation Sequencing of begomovirus genomes. The rapid ability of these viruses to evolve threatens many crops and underscores the importance of using next generation sequencing efficiently to detect and understand the diversity of these viruses. We combined enhanced rolling circle amplification (RCA) with EquiPhi29 polymerase and size selection to generate a cost-effective, short-read sequencing method. This optimized protocol produced short-read sequencing with at least 50% of the reads mapping to the viral reference genome. We provide other insights into common misconceptions about RCA and lessons we have learned from sequencing single-stranded DNA viruses. Our protocol can be used to examine viral DNA as it moves through the entire pathosystem from host to vector, providing valuable information for viral DNA population studies, and would likely work well with other CRESS DNA viruses.HighlightsProtocol for short-read, high throughput sequencing of single-stranded DNA viruses using random primersComparison of the sequencing of total DNA versus size-selected DNAComparison of phi29 and Equiphi29 DNA polymerases for rolling circle amplification of viral single-stranded DNA genomes

CRISPR-Cas12a enhanced rolling circle amplification method for ultrasensitive miRNA detection

2020 ◽  
Vol 158 ◽  
pp. 105239
Author(s):  
Gong Zhang ◽  
Lei Zhang ◽  
Jingtao Tong ◽  
Xianxian Zhao ◽  
Jianlin Ren
Keyword(s):  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
Amplification Method ◽  
Mirna Detection

A T7 exonuclease-assisted cyclic enzymatic amplification method coupled with rolling circle amplification: a dual-amplification strategy for sensitive and selective microRNA detection

2014 ◽  
Vol 50 (13) ◽  
pp. 1576-1578 ◽  
Author(s):  
Liang Cui ◽  
Zhi Zhu ◽  
Ninghang Lin ◽  
Huimin Zhang ◽  
Zhichao Guan ◽  
...  
Keyword(s):  
Rolling Circle Amplification ◽  
Ultrasensitive Detection ◽  
Rolling Circle ◽  
Enzymatic Amplification ◽  
Amplification Method ◽  
Microrna Detection ◽  
Dual Amplification ◽  
Amplification Strategy ◽  
Excellent Selectivity ◽  
T7 Exonuclease

A T7 exonuclease-assisted cyclic enzymatic amplification method (CEAM) was combined with rolling circle amplification (RCA) to develop a RCA–CEAM dual amplification method for ultrasensitive detection of microRNA with excellent selectivity.

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