Sep 22, 2019
Plasmid Isolation: Miniprep (Protocol for GeneJET Plasmid Miniprep Kit)
- 1Wageningen University
- iGEM Wageningen 2019
Protocol Citation: Alba Balletbó 2019. Plasmid Isolation: Miniprep (Protocol for GeneJET Plasmid Miniprep Kit). protocols.io https://dx.doi.org/10.17504/protocols.io.7j4hkqw
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: Working
We use this protocol and it's working
Created: September 22, 2019
Last Modified: September 22, 2019
Protocol Integer ID: 27996
Materials
MATERIALS
GeneJET Plasmid Miniprep KitThermo FisherCatalog #K0502
Centrifuge overnight culture at 4700rpm for 5 minutes and discard the supernatant.
Add 250 µL Resuspension solution and resuspend the cells in this. Transfer the mix to an Eppendorf tube.
Add 250 µL Lysis solution and invert the tube 4-6 times.
Add 350 µL Neutralization solution and invert the tube 4-6 times.
Centrifuge for 5 minutes and transfer the supernatant to a GeneJET spin column tube.
Centrifuge for 1 minute. Empty the flowthrough and place the column back in the collection tube.
Add 500 µL Wash buffer and centrifuge for 1 minute. Discard the flowthrough and place the column back.
Repeat step 7.
Centrifuge for an additional minute to remove all residual ethanol. Place the column in a fresh Eppendorf.
Add 25-50 µL of either Elution buffer or MilliQ to the column and incubate at room temperature for 1 minute.
Centrifuge the column for 1 minute.
Discard the column and store the purified DNA in -20ºC.