scholarly journals Whole genome amplification and exome sequencing of archived schistosome miracidia

Parasitology ◽  
2018 ◽  
Vol 145 (13) ◽  
pp. 1739-1747 ◽  
Author(s):  
Winka Le Clec'h ◽  
Frédéric D. Chevalier ◽  
Marina McDew-White ◽  
Fiona Allan ◽  
Bonnie L. Webster ◽  
...  

AbstractAdult schistosomes live in the blood vessels and cannot easily be sampled from humans, so archived miracidia larvae hatched from eggs expelled in feces or urine are commonly used for population genetic studies. Large collections of archived miracidia on FTA cards are now available through the Schistosomiasis Collection at the Natural History Museum (SCAN). Here we describe protocols for whole genome amplification of Schistosoma mansoni and Schistosome haematobium miracidia from these cards, as well as real time PCR quantification of amplified schistosome DNA. We used microgram quantities of DNA obtained for exome capture and sequencing of single miracidia, generating dense polymorphism data across the exome. These methods will facilitate the transition from population genetics, using limited numbers of markers to population genomics using genome-wide marker information, maximising the value of collections such as SCAN.

2005 ◽  
Vol 26 (3) ◽  
pp. 710-715 ◽  
Author(s):  
Mladen V. Tzvetkov ◽  
Christian Becker ◽  
Bettina Kulle ◽  
Peter Nürnberg ◽  
Jürgen Brockmöller ◽  
...  

2007 ◽  
Vol 2 (8) ◽  
pp. S442-S443
Author(s):  
Trevor J. Pugh ◽  
Allen D. Delaney ◽  
Stephane Flibotte ◽  
Noushin Farnoud ◽  
Irene Li ◽  
...  

PLoS ONE ◽  
2018 ◽  
Vol 13 (3) ◽  
pp. e0193689 ◽  
Author(s):  
Alberto Ferrarini ◽  
Claudio Forcato ◽  
Genny Buson ◽  
Paola Tononi ◽  
Valentina del Monaco ◽  
...  

Parasitology ◽  
2009 ◽  
Vol 137 (1) ◽  
pp. 27-36 ◽  
Author(s):  
MAHA BOUZID ◽  
DARREN HEAVENS ◽  
KRISTIN ELWIN ◽  
RACHEL M. CHALMERS ◽  
STEPHEN J. HADFIELD ◽  
...  

SUMMARYClinical and environmental isolates of pathogens are often unique and may be unculturable, yielding a very limited amount of DNA for genetic studies.Cryptosporidiumin particular are difficult to propagate. Whole genome amplification (WGA) is a valuable technique for amplifying genomic material. In this study, we tested 5 WGA commercial kits usingCryptosporidiumclinical isolates. DNA of 5C. hominisand 5C. parvumclinical isolates andC. parvumIOWA reference strain were used. The majority of the samples were amplified by all of the kits tested. The integrity and fidelity of the amplified genomic DNA were assessed by sequence analysis of several PCR products of varying length. We found evidence that one kit in particular may be more error prone while another seemed the more suitable kit forCryptosporidiumclinical samples, generating high molecular weight DNA from all the samples with high fidelity. Thus WGA was found to be a useful technique for producing amplified DNA suitable for downstream genotyping techniques and archiving ofCryptosporidiumclinical isolates.


2020 ◽  
Author(s):  
Veronica Gonzalez ◽  
Sivaraman Natarajan ◽  
Yuntao Xia ◽  
David Klein ◽  
Robert Carter ◽  
...  

AbstractImprovements in whole genome amplification (WGA) would enable new types of basic and applied biomedical research, including studies of intratissue genetic diversity that require more accurate single-cell genotyping. Here we present primary template-directed amplification (PTA), a new isothermal WGA method that reproducibly captures >95% of the genomes of single cells in a more uniform and accurate manner than existing approaches, resulting in significantly improved variant calling sensitivity and precision. To illustrate the new types of studies that are enabled by PTA, we developed direct measurement of environmental mutagenicity (DMEM), a new tool for mapping genome-wide interactions of mutagens with single living human cells at base pair resolution. In addition, we utilized PTA for genome-wide off-target indel and structural variant detection in cells that had undergone CRISPR-mediated genome editing, establishing the feasibility for performing single-cell evaluations of biopsies from edited tissues. The improved precision and accuracy of variant detection with PTA overcomes the current limitations of accurate whole genome amplification, which is the major obstacle to studying genetic diversity and evolution at cellular resolution.


RSC Advances ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 4617-4621
Author(s):  
Jing Tu ◽  
Yi Qiao ◽  
Yuhan Luo ◽  
Naiyun Long ◽  
Zuhong Lu

Monitoring multiple displacement amplification by fluorescence signals.


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