In this video I will demonstrate how to prepare and load a standard DNA ladder. One that does not come and are ready to load format because standard any be DNA ladders are concentrated DNA solutions. They need to be diluted. And a loading die should be added prior to loading onto a gel to prepare your DNA ladder. And your samples you'll need some small plastic tubes, 500 micro liter tubes work. Well, because we often load a lot of samples at once we use a 96-well plate.
You require about 1 micro. Liter of. The prepared DNA ladder mix per millimeter of gel Lane.
Most standard gel lanes are around 5 millimeters wide. So 6, microliters is a good volume to load and easy to prepare. Now we can prepare our ladder, add 4 microliters of water to your tube or well. And then add one microliter of the 6x purple loading die without SDS, which is supplied with most of our DNA ladders mix by flicking the tube.
And spinning briefly, add one microliter of the DNA ladder to the diluted loading dye mixture mix again by. Flicking the tube and spinning briefly to gather the prepared ladder at the bottom of the tube, the DNA ladder is then ready to load on your gel slowly pipette, the six microliters of the prepared DNA ladder mix into the desired well load your prepared samples. And then you are ready to run your gel in the appropriate migration conditions. The DNA ladder mix can be scaled up.
If you need to load multiple gel lanes, simply prepare, a 1x dilution of the 6x purple loading dye. No SDS in advance by diluting. It with water for each gel Lane that you'd like to run add 5, microliters of the 1x purple loading dye dilution into a micro fiche tube, then add one microliter of the DNA ladder, flick the tube with your finger and spin it briefly again. The DNA ladder is then ready to load.
You want to avoid diluting the DNA ladder and water directly without the loading dye present. When the ladder is in the presence of water without loading dye. Some DNA fragments can denature. This is why it is important to.
Add the ladder to the diluted loading dye mixture alternatively. You can use the buffer instead of water for any DNA ladder, dilution to prevent DNA denaturation for your convenience. Any b also offers an array of DNA ladders available and ready to load formats. These ladders are really easy to use invert. The vial a couple of times to mix then just load directly onto your gel that's it. If you are ever unsure of how to use any of our products you can always contact our technical support scientists by.
Phone or by emailing info at any be com, we're always happy to help.