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URL: https://dx.doi.org/DOI:10.17504/protocols.io.pvgdn3w

Authors: Isabelle Vea, Andrés G. de la Filia, Stevie Bain

Group: Mealybug Team

Summary: Protocol to extract DNA from female mealybug (P.citri) for genomic sequencing.This protocol was established by Stevie Bain and Andres Garcia de la Filia in the Ross lab (University of Edinburgh, IEB), which includes a combination of two kits. I modified it slightly based on Dom Laetsch protocol version 1.1 (Blaxter lab, University of Edinburgh, IEB)Main modifications:13,000 rpm centrifuge at 4deg all the time instead of 13000 rpm at room temperatureAmounts of reagents kept from Andres's protocol but use Cell lysis buffer and Protein precitpitation solution from Qiagen (Dom's)Overnight incubation at 56 is in a heated chambre with a rocking platform to allow gentle mixing All vortexing steps were changed to mixing by inversion and rotation as in Dom's protocol.Beware:RNase A concentration should be 100mg/mL the first time and if there is still RNA left, do a second step of RNase A at 20ug/mL as final concentration

Affiliations: University of Illinois at Chicago, University of Edinburgh, University of Edinburgh, University of Edinburgh

Version: 1

Publication Date: 2021

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