In Situ Detection of Messenger RNA Using Digoxigenin-Labeled Oligonucleotides and Rolling Circle Amplification

2001 ◽  
Vol 70 (3) ◽  
pp. 281-288 ◽  
Author(s):  
Yi Zhou ◽  
Margaret Calciano ◽  
Stefan Hamann ◽  
J.H. Leamon ◽  
Tod Strugnell ◽  
...  
Keyword(s):  
Messenger Rna ◽  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
In Situ Detection

scholarly journals In situ Detection of Specific DNA Double Strand Breaks using Rolling Circle Amplification

Cell Cycle ◽  
2005 ◽  
Vol 4 (12) ◽  
pp. 1767-1773 ◽  
Author(s):  
Jia Li ◽  
C.S.H. Young ◽  
Paul M. Lizardia ◽  
David F. Stern
Keyword(s):  
Rolling Circle Amplification ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Rolling Circle ◽  
Strand Breaks ◽  
In Situ Detection

scholarly journals Detection of Low-Copy-Number Genomic DNA Sequences in Individual Bacterial Cells by Using Peptide Nucleic Acid-Assisted Rolling-Circle Amplification and Fluorescence In Situ Hybridization

2007 ◽  
Vol 73 (7) ◽  
pp. 2324-2328 ◽  
Author(s):  
Irina Smolina ◽  
Charles Lee ◽  
Maxim Frank-Kamenetskii
Keyword(s):  
Dna Sequences ◽  
Peptide Nucleic Acid ◽  
Rolling Circle Amplification ◽  
Bacterial Genome ◽  
Bacterial Cells ◽  
Rolling Circle ◽  
Low Copy Number ◽  
Short Signature ◽  
In Situ Detection

ABSTRACT An approach is proposed for in situ detection of short signature DNA sequences present in single copies per bacterial genome. The site is locally opened by peptide nucleic acids, and a circular oligonucleotide is assembled. The amplicon generated by rolling circle amplification is detected by hybridization with fluorescently labeled decorator probes.

Toehold-initiated Rolling Circle Amplification for Visualizing Individual MicroRNAs In Situ in Single Cells

2014 ◽  
Vol 126 (9) ◽  
pp. 2421-2425 ◽  
Author(s):  
Ruijie Deng ◽  
Longhua Tang ◽  
Qianqian Tian ◽  
Ying Wang ◽  
Lei Lin ◽  
...  
Keyword(s):  
Single Cells ◽  
Rolling Circle Amplification ◽  
Rolling Circle

scholarly journals Visualization and Enumeration of Bacteria Carrying a Specific Gene Sequence by In Situ Rolling Circle Amplification

2005 ◽  
Vol 71 (12) ◽  
pp. 7933-7940 ◽  
Author(s):  
Fumito Maruyama ◽  
Takehiko Kenzaka ◽  
Nobuyasu Yamaguchi ◽  
Katsuji Tani ◽  
Masao Nasu
Keyword(s):  
Gene Sequence ◽  
Rolling Circle Amplification ◽  
Specific Gene ◽  
Toxin Gene ◽  
Dna Uptake ◽  
Fluorescent Intensity ◽  
Prolonged Incubation ◽  
Rolling Circle ◽  
E Coli

ABSTRACT Rolling circle amplification (RCA) generates large single-stranded and tandem repeats of target DNA as amplicons. This technique was applied to in situ nucleic acid amplification (in situ RCA) to visualize and count single Escherichia coli cells carrying a specific gene sequence. The method features (i) one short target sequence (35 to 39 bp) that allows specific detection; (ii) maintaining constant fluorescent intensity of positive cells permeabilized extensively after amplicon detection by fluorescence in situ hybridization, which facilitates the detection of target bacteria in various physiological states; and (iii) reliable enumeration of target bacteria by concentration on a gelatin-coated membrane filter. To test our approach, the presence of the following genes were visualized by in situ RCA: green fluorescent protein gene, the ampicillin resistance gene and the replication origin region on multicopy pUC19 plasmid, as well as the single-copy Shiga-like toxin gene on chromosomes inside E. coli cells. Fluorescent antibody staining after in situ RCA also simultaneously identified cells harboring target genes and determined the specificity of in situ RCA. E. coli cells in a nonculturable state from a prolonged incubation were periodically sampled and used for plasmid uptake study. The numbers of cells taking up plasmids determined by in situ RCA was up to 106-fold higher than that measured by selective plating. In addition, in situ RCA allowed the detection of cells taking up plasmids even when colony-forming cells were not detected during the incubation period. By optimizing the cell permeabilization condition for in situ RCA, this method can become a valuable tool for studying free DNA uptake, especially in nonculturable bacteria.

Lectin-mediated in situ rolling circle amplification on exosomes for probing cancer-related glycan pattern

2018 ◽  
Vol 1039 ◽  
pp. 108-115 ◽  
Author(s):  
Yimei Feng ◽  
Yuna Guo ◽  
Yiran Li ◽  
Jing Tao ◽  
Lin Ding ◽  
...  
Keyword(s):  
Rolling Circle Amplification ◽  
Rolling Circle ◽  
Glycan Pattern

scholarly journals In situ detection of non-polyadenylated RNA molecules using Turtle Probes and target primed rolling circle PRINS

2007 ◽  
Vol 7 (1) ◽  
pp. 69 ◽  
Author(s):  
Magnus Stougaard ◽  
Jakob S Lohmann ◽  
Magdalena Zajac ◽  
Stephen Hamilton-Dutoit ◽  
Jørn Koch
Keyword(s):  
Rolling Circle ◽  
Rna Molecules ◽  
Polyadenylated Rna ◽  
In Situ Detection
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