Automated, high-throughput DNA extraction protocol for disposable label free, microfluidics integrating DNA biosensor for oil palm pathogen, Ganoderma boninense

2020 ◽  
Vol 92 ◽  
pp. 447-456 ◽  
Author(s):  
Adilah Ayoib ◽  
Uda Hashim ◽  
Subash C.B. Gopinath
2018 ◽  
Vol 24 (2) ◽  
pp. 77-86
Author(s):  
Sri Wening ◽  
Agus Eko Prasetyo ◽  
Tjut Ahmad Perdana Rozziansha ◽  
Agus Susanto

African pollination weevil (Elaeidobius kamerunicus Faust) has an important role in the productivity of Indonesian oil palm plantation. Up to now, there has not been a comprehensive biological study of the species at molecular level. The basic knowledge is very useful for exploitation of the weevil for effective oil palm fruit set development. This research aimed to obtain DNA extraction protocol of E. kamerunicus for DNA fingerprinting of the species. Results showed that using a DNA extraction kit,material disruption by using micro pestle resulted the highest quantity of DNA, while there were no significant differences of resulted DNA quantity among treatments using tissue lyser for material disruption. DNA extracted by using micro pestle or tissue lyser for material disruption is adequate for DNA fingerprinting using AFLP (Amplified Fragment Length Polymorphism) and sequencing techniques.


2020 ◽  
Author(s):  
santiago vilanova ◽  
David Alonso ◽  
Pietro Gramazio ◽  
Mariola Plazas ◽  
Edgar Garcia Fortea ◽  
...  

Abstract Background The use of sequencing and genotyping platforms has undergone dramatic improvements, enabling the generation of a wealth of genomic information. Despite this progress, the availability of high-quality genomic DNA (gDNA) in sufficient concentrations is often a main limitation, especially for third-generation sequencing platforms. A variety of DNA extraction methods and commercial kits are available. However, many of these are costly and frequently give either low yield or low-quality DNA, inappropriate for next generation sequencing (NGS) platforms. Here, we describe a fast and inexpensive DNA extraction method (SILEX) applicable to a wide range of plant species and tissues. Results SILEX is a high-throughput DNA extraction protocol, based on the standard CTAB method with a DNA silica matrix recovery, which allows obtaining NGS-quality high molecular weight genomic plant DNA free of inhibitory compounds. SILEX was compared with a standard CTAB extraction protocol and a common commercial extraction kit in a variety of species, including recalcitrant ones, from different families. In comparison with the other methods, SILEX yielded DNA in higher concentrations and of higher quality. Manual extraction of 48 samples can be done in 96 min by one person at a cost of 0.12 €/sample of reagents and consumables. Hundreds of tomato gDNA samples obtained with either SILEX or the commercial kit were successfully genotyped with Single Primer Enrichment Technology (SPET) with the Illumina HiSeq 2500 platform. Furthermore, DNA extracted from Solanum elaeagnifolium using this protocol was assessed by Pulsed-field gel electrophoresis (PFGE), obtaining a suitable size ranges for most sequencing platforms that required high-molecular-weight DNA such as Nanopore or PacBio. Conclusions A high-throughput, fast and inexpensive DNA extraction protocol was developed and validated for a wide variety of plants and tissues. SILEX offers an easy, scalable, efficient and inexpensive way to extract DNA for various next-generation sequencing applications including SPET and Nanopore among others.


Author(s):  
Lisa Marotz ◽  
Tara Schwartz ◽  
Luke Thompson ◽  
Greg Humphrey ◽  
Grant Gogul ◽  
...  

Crop Science ◽  
2002 ◽  
Vol 42 (6) ◽  
pp. 2088-2091 ◽  
Author(s):  
Kristi L. Hill-Ambroz ◽  
Gina L. Brown-Guedira ◽  
John P. Fellers

2003 ◽  
Vol 21 (4) ◽  
pp. 459-460 ◽  
Author(s):  
Emma S. Mace ◽  
Kutokshi K. Buhariwalla ◽  
Hutokshi K. Buhariwalla ◽  
Jonathan H. Crouch

Aquaculture ◽  
2006 ◽  
Vol 255 (1-4) ◽  
pp. 597-599 ◽  
Author(s):  
Noam Zilberman ◽  
Sharon Reikhav ◽  
Gideon Hulata ◽  
Micha Ron

Sign in / Sign up

Export Citation Format

Share Document