scholarly journals Purification of High Molecular Weight Genomic DNA from Powdery Mildew for Long-Read Sequencing

10.3791/55463 ◽  
2017 ◽  
Author(s):  
Joanna M. Feehan ◽  
Katherine E. Scheibel ◽  
Salim Bourras ◽  
William Underwood ◽  
Beat Keller ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Powdery Mildew ◽  
High Molecular Weight ◽  
Genomic Dna ◽  
Long Read

scholarly journals Extraction of high-molecular-weight genomic DNA for long-read sequencing of single molecules

BioTechniques ◽  
2016 ◽  
Vol 61 (4) ◽  
Author(s):  
Baptiste Mayjonade ◽  
Jérôme Gouzy ◽  
Cécile Donnadieu ◽  
Nicolas Pouilly ◽  
William Marande ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Genomic Dna ◽  
Single Molecules ◽  
Long Read

scholarly journals Extraction of high-molecular-weight genomic DNA for long-read sequencing of single molecules

BioTechniques ◽  
2017 ◽  
Vol 62 (1) ◽  
Author(s):  
Baptiste Mayjonade ◽  
Jérôme Gouzy ◽  
Cécile Donnadieu ◽  
Nicolas Pouilly ◽  
William Marande ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Genomic Dna ◽  
Single Molecules ◽  
Long Read

scholarly journals Optimization of high molecular weight DNA extraction methods in shrimp for a long-read sequencing platform

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e10340
Author(s):  
Pacharaporn Angthong ◽  
Tanaporn Uengwetwanit ◽  
Wirulda Pootakham ◽  
Kanchana Sittikankaew ◽  
Chutima Sonthirod ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Genomic Dna ◽  
Extraction Methods ◽  
Marine Organisms ◽  
High Quality ◽  
Sequencing Platform ◽  
Global Food Security ◽  
Long Read ◽  
Ctab Method ◽  
Global Food

Marine organisms are important to global food security as they are the largest source of animal proteins feeding mankind. Genomics-assisted aquaculture can increase yield while preserving the environment to ensure sufficient and sustainable production for global food security. However, only few high-quality genome sequences of marine organisms, especially shellfish, are available to the public partly because of the difficulty in the sequence assembly due to the complex nature of their genomes. A key step for a successful genome sequencing is the preparation of high-quality high molecular weight (HMW) genomic DNA. This study evaluated the effectiveness of five DNA extraction protocols (CTAB, Genomic-tip, Mollusc DNA, TIANamp Marine Animals DNA, and Sbeadex livestock kits) in obtaining shrimp HMW DNA for a long-read sequencing platform. DNA samples were assessed for quality and quantity using a Qubit fluorometer, NanoDrop spectrophotometer and pulsed-field gel electrophoresis. Among the five extraction methods examined without further optimization, the Genomic-tip kit yielded genomic DNA with the highest quality. However, further modifications of these established protocols might yield even better DNA quality and quantity. To further investigate whether the obtained genomic DNA could be used in a long-read sequencing application, DNA samples from the top three extraction methods (CTAB method, Genomic-tip and Mollusc DNA kits) were used for Pacific Biosciences (PacBio) library construction and sequencing. Genomic DNA obtained from Genomic-tip and Mollusc DNA kits allowed successful library construction, while the DNA obtained from the CTAB method did not. Genomic DNA isolated using the Genomic-tip kit yielded a higher number of long reads (N50 of 14.57 Kb) than those obtained from Mollusc DNA kits (N50 of 9.74 Kb). Thus, this study identified an effective extraction method for high-quality HMW genomic DNA of shrimp that can be applied to other marine organisms for a long-read sequencing platform.

Extracting high molecular weight genomic DNA from Saccharomyces cerevisiae

2018 ◽  
Author(s):  
Erwan DENIS ◽  
Erwan Denis ◽  
Sophie Sanchez ◽  
Barbara Mairey ◽  
Odette Beluche ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Molecular Weight ◽  
High Molecular Weight ◽  
Genomic Dna

Calcium Phosphate-Mediated Transfection of Cells with High-Molecular-Weight Genomic DNA

2019 ◽  
Vol 2019 (10) ◽  
pp. pdb.prot095448 ◽  
Author(s):  
Priti Kumar ◽  
Arvindhan Nagarajan ◽  
Pradeep D. Uchil
Keyword(s):  
Molecular Weight ◽  
Calcium Phosphate ◽  
High Molecular Weight ◽  
Genomic Dna
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