Apr 02, 2022

Public workspaceeDNA extraction with kaneka V.1

DOI
dx.doi.org/10.17504/protocols.io.dm6gpb9xplzp/v1
  • Fei Xia1
  • 1Keio University
xiafei
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DOI: dx.doi.org/10.17504/protocols.io.dm6gpb9xplzp/v1
External link: https://onlinelibrary.wiley.com/doi/full/10.1111/1755-0998.13448
Protocol Citation: Fei Xia 2022. eDNA extraction with kaneka. protocols.io https://dx.doi.org/10.17504/protocols.io.dm6gpb9xplzp/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: In development
We are still developing and optimizing this protocol
Created: April 02, 2022
Last Modified: April 02, 2022
Protocol Integer ID: 60213
Abstract
We used the Kaneka Simple DNA Extraction Kit version 2 (Kaneka) for on-site DNA extraction from Sterivex. We injected 500 µl of Solution A of the kit into the Sterivex and shook it by hand for 1 min. We collected the Solution A buffer in the Sterivex by connecting the syringe with the connecting tube. We added 70 µl of Solution B into the collected Solution A buffer in a 1.5-ml microtube and centrifuged (~2,000 g) with a portable centrifuge for 3 min. We collected 200 µl of the supernatant solution.
Materials
stervix sample
カネカ簡易DNA抽出キット Version 2
1.5ml tube
50ml tube




Inject Amount250 µL of A buffer into the Sterivex, Duration00:01:00 shake and rotate.

1m


Put Sterivex into 50ml tube and secured for centrifugation Centrifigation5000 rpm, 00:05:00
5m

Remove the liquid from the inlet and put into the 1.5ml tube.


Add Amount40 µL of B buffer to the 1.5ml tube.



Mix by vortexing.


Store the samples at Temperature-20 °C .