How to cast and run a Polyacrylamide Gel in the Wave Maxi Vertical

The new VS20 WAVE Maxi System is Cleaver Scientifics latest product innovation for large format vertical gel electrophoresis. It’s designed to perform a variety of separations, including first and second-dimension SDS-PAGE, native, preparative, gradient and high-resolution nucleic acid electrophoresis, plus capillary tube gel IEF and electroblotting and is one of the most versatile maxi vertical systems available.

How to cast and run a Polyacrylamide Gel in the Wave Maxi Vertical

In this guide we’re going to show you how to cast and run polyacrylamide gel in the Wave Maxi Vertical.

What you’ll need

Here’s what you will need:

  • Gel casting and running upstand
  • Casting base with mat
  • 2 x Plain glass plates with 1mm bonded spaces
  • 2 x Notch glass plates
  • 2 x 1mm 24 sample combs

Prepare The Glass Plate Sandwich

Ensure that all components are clean, dry and free of any chips or cracks before making a glass plate sandwich. If running four gels instead of two, position a notched glass plate with bonded spaces over the single gel sandwich. Also, use the thinner yellow clamps in the casting upstand. Unscrew the red and black screws in the upstand to release the locking mechanism, allowing the green gel clamps to sit in the resting slots.

With the green clamps now out of the way, carefully slide the glass plate sandwich into the upstand, lowering the glass plates until they are flush with the bottom of the side cheeks on the lab bench surface, not on the casting base.

Turn the red and black screws.

These will push down the green clamps to secure the gel sandwich against the gasket. The Notched Glass Plates MUST ALWAYS Face Innermost.

Insert the dummy plate if only casting and running one gel. Check that plates are flush with each other.

Pull out the cam pins from the casting base and rotate them so that they point downwards into the lab bench surface.

Transfer Upstand To Casting Base

Transfer the upstand containing glass plates to the casting base, ensuring that the glass plates sit evenly on the ultra soft silicon mat.

It may be necessary to apply gentle pressure to the top of the upstand to enable the cams to locate in the holes within the upstand.

Turn Both Cam Handles 90 Degrees In Opposite Directions.

Secure the upstand to the casting mat by turning the cam handles in opposite directions.

By 90 degrees the cams will feel tight.

Pour the gel

The gel solution is now poured between the glass plates and the combs inserted.

Typical constituents of a denaturing gel include:

  1. Acrylamide
  2. Tris-Glycine
  3. SDS
  4. Distilled Water
  5. APS
  6. TEMED

The standard gel formulation may vary according to the acrylamide percentage, usually higher in a  resolving gel but the constituents remain the same.

Remove the combs from the polymerised gels.

Align colour coded screws with red and black thumb locators in the tank.

Transfer Upstand To The Tank.

If connecting to a chiller, make sure that the cooling coil is in situ before inserting the upstand.

Fill Inner And Outer Chambers With Buffer.

Use the provided loading indicator as a template to position the pipette above each well during loading. Load the protein samples mixed with Laemmli Buffer. After loading, replace the lid and insert the power cables into the corresponding colour coded ports in the power supply.

Power Supply Settings

Typical Power Supply Settings:

  • 100 – 350 Volts (start to end)
  • Constant Current: 35mA
  • Time: 3.5 to 5 Hours

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