Analysis of Multiallelic CNVs by Emulsion Haplotype Fusion PCR

2016 ◽  
pp. 155-165 ◽  
Author(s):  
Jess Tyson ◽  
John A. L. Armour
Keyword(s):  
Fusion Pcr

scholarly journals Efficient One-Step Fusion PCR Based on Dual-Asymmetric Primers and Two-Step Annealing

2017 ◽  
Vol 60 (2) ◽  
pp. 92-99 ◽  
Author(s):  
Yilan Liu ◽  
Jinjin Chen ◽  
Anders Thygesen
Keyword(s):  
Fusion Pcr ◽  
One Step

scholarly journals Inverse Fusion PCR Cloning

PLoS ONE ◽  
2012 ◽  
Vol 7 (4) ◽  
pp. e35407 ◽  
Author(s):  
Markus Spiliotis
Keyword(s):  
Pcr Cloning ◽  
Fusion Pcr

scholarly journals Pcr-Based Construction and Transformation of Cody Gene Deletion Construct in Streptococcus Pneumoniae

2017 ◽  
Vol 16 (1) ◽  
Author(s):  
Aisyah Mohamed Rehan ◽  
Mohammad Izwan Enche Othman ◽  
Nor Munirah Mohd Amin ◽  
Intan Azura Shahdan ◽  
Hanani Ahmad Yusof@Hanafi
Keyword(s):  
Streptococcus Pneumoniae ◽  
Gene Deletion ◽  
Single Gene ◽  
Crystallographic Structure ◽  
Mortality And Morbidity ◽  
Fusion Pcr ◽  
Deletion Construct ◽  
Hyaluronate Lyase ◽  
Pcr Method ◽  
Single Gene Deletion

Introduction: Streptococcus pneumoniae (S. pneumoniae) is a gram-positive diplococci belonging to the genus Streptococcus and it is a well-studied pathogenic bacterium. Pneumococcal diseases such as otitis media, pneumonia, sepsis and meningitis caused by pathogenic strains of S. pneumoniae still brought significant mortality and morbidity worldwide. The pathogenicity of S. pneumoniae is exerted by various virulence factors and one of it is the enzyme hyaluronate lyase. Hyaluronate lyase plays a major role in the invasive capability of S. pneumoniae. Its mechanism of action and crystallographic structure have been determinedbut its regulatory mechanism is still poorly understood. Drawing connections between the nutritional behaviour and invasive property of S. pneumoniae, CodY regulator is hypothesized as a potential hyaluronate lyase regulator. This work was aimed to construct CodY deficient mutant of S. pneumoniae to form foundational work for the study of CodY regulatory effect on hyaluronate lyase. Materials and method: A single gene-deletion method was chosen to create CodY mutant construct containing aphA-3 gene (encoding kanamycin resistance protein) replacement cassette by employing recombinant fusion PCR method. A single band DNA product at the expected size of 2,563 bp after recombinant fusion PCR was visualized on agarose gel electrophoresis. This indicated that the linear recombinant amplicon has been successfully constructed. The recombinant amplicon was transformed into S. pneumoniae cells of Malaysian local clinical isolate by employing the competence stimulating peptide (CSP-1) to enhance and induce natural competence. Results: No viable CodY mutant was evident post-transformation. The negative transformation result was postulated to be due to the essential nature of CodY regulator. Conclusion: This work can provide basis for recombinant fusion PCR method in designing and constructing single gene deletion construct. However, further work is needed to identify the regulatory agent of hyaluronate lyase.

Haplotype-Fusion PCR

2006 ◽  
Author(s):  
Daniel Turner ◽  
Matthew Hurles
Keyword(s):  
Fusion Pcr

Fusion PCR via Novel Overlap Sequences

2012 ◽  
pp. 97-110 ◽  
Author(s):  
Kamonchai Cha-aim ◽  
Hisashi Hoshida ◽  
Tomoaki Fukunaga ◽  
Rinji Akada
Keyword(s):  
Fusion Pcr

scholarly journals Application of fusion PCR to the amplification of full-length ORF sequences of different splicing variants of NuMA1 from HeLa cells

2017 ◽  
Vol 49 (10) ◽  
pp. 962-965
Author(s):  
Jin Wu ◽  
Guanting Lu ◽  
Jianwei Wu ◽  
Hua Yang ◽  
Zhicao Yu ◽  
...  
Keyword(s):  
Hela Cells ◽  
Full Length ◽  
Fusion Pcr ◽  
Splicing Variants

Fusion PCR and gene targeting in Aspergillus nidulans

2006 ◽  
Vol 1 (6) ◽  
pp. 3111-3120 ◽  
Author(s):  
Edyta Szewczyk ◽  
Tania Nayak ◽  
C Elizabeth Oakley ◽  
Heather Edgerton ◽  
Yi Xiong ◽  
...  
Keyword(s):  
Aspergillus Nidulans ◽  
Gene Targeting ◽  
Fusion Pcr

scholarly journals Linking plasmid-based beta-lactamases to their bacterial hosts using single-cell fusion PCR

2021 ◽  
Author(s):  
Peter J. Diebold ◽  
Felicia N. New ◽  
Michael Hovan ◽  
Michael J. Satlin ◽  
Ilana L. Brito
Keyword(s):  
Neutropenic Patient ◽  
Multidrug Resistant ◽  
Metal Resistance ◽  
Alternative Methods ◽  
Rrna Gene ◽  
Host Bacterium ◽  
Environmental Pressures ◽  
Beta Lactamases ◽  
Fusion Pcr ◽  
Long Read

AbstractThe horizonal transfer of plasmid-encoded genes allows bacteria to adapt to constantly shifting environmental pressures, bestowing functional advantages to their bacterial hosts such as antibiotic resistance, metal resistance, virulence factors, and polysaccharide utilization. However, common molecular methods such as short- and long-read sequencing of microbiomes cannot associate extrachromosomal plasmids with the genome of the host bacterium. Alternative methods to link plasmids to host bacteria are either laborious, expensive or prone to contamination. Here we present the One-step Isolation and Lysis PCR (OIL-PCR) method, which molecularly links target ARGs with the bacterial 16S rRNA gene via fusion PCR performed within an emulsion. After validating this method, we apply it to identify the bacterial hosts of three clinically relevant beta-lactamases in a neutropenic patient population who are particularly vulnerable multidrug-resistant infections. We detect novel associations of two low-abundance genera, Romboutsia and Agathobacter, with a multi-drug resistant plasmid harbored by Klebsiella pneumoniae. We put forth a robust, accessible, and high-throughput platform for sensitively surveying the bacterial hosts of mobile genes in complex microbial communities.

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