Magnetic bead purification throughput boost with the VIAFLO 96/384 electronic pipette (video)
Magnetic bead purification throughput boost with the VIAFLO 96/384 electronic pipette
Hi everyone, in this video we are going to show you, how the VIAFLO 96 384 electronic pipette can make DNA purification with magnetic beads faster, less error-prone and more reproducible. DNA purification with magnetic beads involves bringing samples and beads together, to begin the DNA binding process. After washing away the impurities the DNA saluted from the beads and is then ready to be used.
To start a process of using the VIAFLO 96 electronic pipette together with magnetic beads, you need to have the following ready. Magnetic beads of course, for this protocol we are using MPI X 3 beads. Samples for this protocol, we have got a 96 by a plate filled in each well with 10 microliters samples to be purified. Magnets this 96-well ring magnet has a standard plate footprint, which allows it to be used in a VIAFLO 96. 70% ethanol as well as the elution buffer and naturally, we'll be using the VIAFLO 96 384 base unit together with a 96 channel head with a volume range of 5 to 125 micro litre.
In this particular protocol we'll be using this repositioning stage. We will be working with low attentions they arrived a little tips. The steps needed to run this procedure are saved as one single custom program called MPI X p.m. on the VIAFLO 96. It contains all the necessary volumes prompts and Heights.
To start the magnetic beads need to be transferred into 12 PCR tubes using a single channel - a 50 micro litre VIAFLO electronic pipette and the repeat dispense program adding 154 micro liter per tube. Afterwards the 50 micro litre 12 channel VIAFLO pipette can be used to transfer 18 microliter of the beads from the PCR tubes into a 96-well plate, which has already been filled with time microliter of the desired samples.
Now we are ready to transfer the sample play to the VIAFLO 96, to begin the purification process. The setup should look like this, the left position or A should have drip tips. The middle position or AB has the 96-well plate with beats loaded. And the right position or B as the magnet. The screen tells the user what the next step is. So all that's needed, is the user to move the instrument to the lab wear.
There's no chance of crashing since the MP u XP program has already defined what the bottom of the specific lab wears and the tips are always guided into correct position. First the beads and samples are mixed thoroughly to ensure a homogeneous mix and left to incubate for five minutes for binding.
Afterwards the sample plate is transferred to the magnet on position B and given two minutes to allow the beads to separate from the solution. Now the supernatant is collected using a very slow speed. The height of the aspiration, like all others in the protocol, is already defined in order to prevent bit carryover and avoid wasting sample. After placing the automation frankly reservoir on position AB the supernatant can be dispensed into it, for waste.
Next, the washing steps can begin. Before starting, fresh tips are electronically loaded after the used ones are ejected. The tip break is removed and in its place the reservoir containing 70% ethanol is added. Before we begin pipetting with ethanol, it is important to be wet the tip with ethanol first, to help prevent dripping tips since ethanol can be tricky to pipe it. Here, the low retention tips really help as well. Once pre-wetting is finished, the three positioning stage, the slid to the left and the ethanol, is then transferred to the samples.
After an incubation period of 30 seconds, the supernatant is once again removed and as before, the perfect height is already exactly defined. It is then discarded in the waste other one. This washing procedure is then repeated again, using new tips. After the second washing, it is especially important, to remove all ethanol from the sample before continuing.
With the washing finished, sample Lucien can now begin. The sample plate is removed from the magnet and placed on position AB. And after loading new tips the elution buffer in the reservoir is placed on position A. 40 microliter of dilution buffer is then added to the sample plate and mixed together, so early in order to homogeneously resuspend the beads.
Afterwards the plate is allowed to incubate for two minutes in order for the DNA to unbind from the beads. Then the sava plate is placed back on the magnet for one minute, so that the beads can separate. As this is happening in empty 96-well dilution plate for the purified DNA sample is placed on position AB and mutants are loaded one last time. With these new tips 40 microliter of dill weight is collected from the sample plate, with the correct height pre-programmed to eliminate B carryover. Enter Spence into the target illusion plate.
Now we are finished. 96 samples of the purified DNA ready for further processing. For those, who prefer level of automation the VIAFLO 96 can also be run in automatic mode, which further minimize the interactions of the user from the purification protocol. This is especially useful in tight spaces for example, if being run under laminar flow.
In addition, if you're not working with 96 samples at one time, the VIAFLO 96 is able to also work with less tips loaded. Giving the ability to purify a smaller number of samples. The entire pipetting protocol is available for download from our website.