May 21, 2024
MagMAX™ CORE Nucleic Acid Purification Kit
This protocol is a draft, published without a DOI.
- 1Moredun Research Institute
Protocol Citation: Karen Keegan 2024. MagMAX™ CORE Nucleic Acid Purification Kit. protocols.io https://protocols.io/view/magmax-core-nucleic-acid-purification-kit-dd3m28k6
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: May 21, 2024
Last Modified: May 21, 2024
Protocol Integer ID: 100173
Abstract
MagMAX‱ CORE Nucleic Acid Purification Kit
For step 3 in box above, follow bacterial DNA purification centrifuge step.
For step 2 in box above, add 200 μl of supernatant from step 1 to the Lysis solution
Select
the programme ‘MagMAXCORE_KF96_kf2 on the MagMAX machine (I used MagMAX express 96 particle processor machine)
As per the table above, first get your plates ready before you load them onto the machine. You will need: 2 deep well plates for the wash solutions. LABEL THE PLATES on the side so you know which plate is which and then add in 500 μl of the appropriate wash buffer solution to the amount of columns you will be using (8 samples = one column etc. make sure to fill up a whole column as this is preferable to avoid damaging the machine). If you don’t have enough samples to make up a full column, fill the empty wells with water. The elution plate is a normal (shallow plate). Make sure to label this on the side too and add 90ul of elution buffer to the amount of columns you will be using.
Note that the ‘tip comb’ plate 5 is the empty tip combs placed in an empty standard (shallow) plate. This is placed into the machine first (the screen on the machine tells you which plate to load in – starts on plate 5 and works backwards). When loading in your plates, make sure the A1 of the plate lines up with the A1 indicated in the plate holder of the machine. When placed in, the plates can ‘bounce’ when pressed in lightly. There is no clicking sound or fast securing of the plates in the machine so no need to use force. Close the door of the machine each time you’ve loaded in your plate then press ‘start’ and the machine will move the rotator around so you can place your next plate in (which will also be displayed on the machines screen). Remember you add the plates in the following order: plate 5,4,3,2,1, sample plate.
You will also need to make up your ‘sample’ plate, which is the plate to be added into the machine last. You will need to make a PK/bead mix to go into this sample plate. Make this by:
Set up the sample plate (in a deep well plate) as indicated below:
Once all your plates are loaded in, press start and the machine will immediately start up – you will notice that the machine will grab the tip combs from the plate five position and then swing back to begin working on plate one (your sample plate) and move from plate one to plate four (elution plate). The run takes less ~1 hour. When the run is complete the machine screen will just say ‘paused’. Press stop and then remove all plates by pressing the left and right arrows (or the start button). The elution plate wont have a cover on it so you may wish to place a seal or some parafilm over it.
Additional bead clean up step to remove potential trace magnetic beads leftover from machine - place the elution plate on a magnetic stand for 2 minutes and then take up the eluate from the centre (as beads pulled to the side of the well). The elution volume for the MagMAX is 90 μl but I got back 75-80 μl after bead clean up.
Store at -70/-80°C until further use.