Using Pall’s Vacuum Manifold with Multi-Well Filter Plates Appendix 6.3

Vacuum Manifold for Use with Multi-Well Filter Plates

The multi-well plate vacuum manifold is an anodized aluminum manifold that has been designed and optimized for the vacuum filtration of the AcroPrep® and AcroWell™ lines of multi-well filter plates. The vacuum manifold comes complete with the necessary O-ring and gasket. The control block includes the vacuum pressure gauge, vacuum metering valve, vacuum release valve, and the 1/4 inch hose barb for vacuum line attachment. Included with the vacuum manifold unit is a Delrin spacer block designed to accommodate standard 350 μL receiver plates. The spacer block has been optimized to reduce the space between the receiver plate and the filter plate during vacuum filtration. An accessory kit that contains an additional O-ring, gasket, and allen wrench is also included with the vacuum manifold unit.   

Table 6.1

Specifications for the Multi-Well Plate Vacuum Manifold

Specification	Parameter
Materials of Construction      Vacuum Manifold      Gasket      O-ring      Spacer blocks	Anodized aluminum EPDM (Ethylene propylene) Silicone Delrin
Dimensions      Length      Width      Height      Weight	17.48 cm (6.88 in) 12.37 cm (4.87 in) 8.05 cm (3.17 in) 6.27 lbs
Maximum Operating Vacuum	71.12 cm Hg (28 in. Hg).

Tip: The multi-well plate vacuum manifold can be used with multi-well filter plates that meet the specifications set forth by the Society for Biomolecular Screening (SBS).

Protocol for Vacuum Manifold for Use with Multi-Well Filter Plates

A. Set the Vacuum Manifold to a Desired Negative Pressure Setting Prior to Filtration

1. Switch the on/off valve to the off position (see Figure 6.3). 
2. Attach the vacuum line to the 1/4 inch hose barb.
3. Place a receiver plate on top of the vacuum manifold.
4. Turn on the vacuum pressure.
5. Adjust the negative pressure to the desired setting by adjusting the metering valve.
6. Turn off the vacuum pressure and remove the receiver plate. The vacuum manifold is now set at the desired setting.

B. Filter Using the AcroPrep® or AcroWell™ Filter Plates

1. If collecting the filtrate, remove the top chamber of the vacuum manifold. The indentations on the sides of the manifold allow for the top portion of the manifold to be easily removed. If collecting the retentate, skip to Step 3.
2. Place the appropriate spacer block into the lower chamber of the vacuum manifold. The vacuum manifold can be used with any size receiver plate.
   1. 350 μL receiver plates: to use a 350 μL receiver plate, place the large spacer block (PN 5015) into the lower chamber of the vacuum manifold. Place the 350 μL receiver plate on top of the spacer block.
   2. 1 mL receiver plates: to use 1 mL receiver plates, place the small spacer block (PN 5014) into the lower chamber of the vacuum manifold. Place the 1 mL receiver plate on top of the spacer block. (You can also use a standard 350 μL receiver plate in plate of the 1 mL spacer block.)
   3. 2 mL receiver plates: to use 2 mL receiver plates, place the 2 mL receiver plate directly into the lower chamber of the vacuum manifold.
3. Before replacing the top chamber of the vacuum manifold, ensure that all surfaces are free from dirt, debris, and any particulate matter that may have accumulated on the vacuum manifold, O-ring, and gasket.
   Tip: If the O-ring and gasket are not clean, you will not obtain a proper seal.
4. Replace the top chamber of the vacuum manifold.
5. Place the filter plate on the gasket located on the top chamber of the vacuum manifold. Ensure that the gasket is clean.
6. Ensure that the on/off switch is in the Off position (see Figure 6.3).
7. Connect the vacuum line to the 1/4 inch hose barb.
8. When ready to evacuate the filter plate, turn the on/off switch on the vacuum manifold to the On position. You may need to press lightly on the filter plate to engage the vacuum seal.
9. The wells will begin to evacuate/empty once the vacuum has been applied to the chamber.
10. If you need to adjust the vacuum pressure up or down, adjust by moving the metering valve located to the right of the vacuum pressure gauge.
11. When all of the wells have completely evacuated, turn the on/off valve to the Off position. To release the residual vacuum pressure that remains in the chamber, push the release valve located to the left of the vacuum gauge. The release valve will allow the pressure within the manifold chamber to return to atmospheric pressure and reduce the potential for cross-contamination and spraying of the filtrate. Do not release the vacuum by pulling the corner of the plate as it will degrade the manifold gasket. You can also tap the top of the filter plate prior to removing it to release any hanging drops that may be on attached to the outlet tips.
12. Remove the filter plate and place it aside for further processing or dispose of it properly.
13. If collecting the filtrate, remove the top chamber of the vacuum manifold from the lower portion of the vacuum manifold. 
14. Remove the receiver plate from the lower chamber of the vacuum manifold and utilize the filtrate for further processing.

C. Replace the O-ring

1. Remove existing O-ring from the bottom of the upper chamber.
2. Ensure that the O-ring is free from dirt, debris, and particulate matter.
3. Place new O-ring into place.

D. Replace the Gasket

1. With the allen wrench provided, remove the 12 screws located on the bottom portion of the upper chamber. 
2. Separate the top ring from the bottom section of the upper chamber.
3. Remove the old gasket and clean the gasket pocket area of any dirt and debris.
4. Place the new gasket into the gasket pocket. Place the top ring back onto the bottom section of the upper chamber. Ensure that the alignment posts on the bottom section fit into the alignment holes in the top ring.
5. Invert the upper chamber and replace the 12 screws.
6. Lightly tighten the four corner screws with the allen wrench, then tighten all 12 screws. Once this is completed, recheck the screws with the allen wrench to ensure that all screws are tightly secured.
7. The manifold is now ready for use.

Figure 6.3

Figure 6.4

Figure 6.5

Complementary Products for Vacuum Manifold for Use with Multi-Well Filter Plates

AcroWell™ 96-well filter plates with BioTrace™ NT and BioTrace PVDF membranes exhibit high binding capacities for proteins and nucleic acids.

AcroPrep® 96 and 384-well filter plates are an excellent platform for a wide variety of molecular biology, analytical, and high throughput sample preparation and detection applications.

Ordering Information for Purification on an Acrodisc® Unit with Mustang® Q Membrane

Part Number	Description	Pkg
5017	Multi-well Plate Vacuum Manifold	1/pkg
5014	1 mL Receiver Plate Spacer Block	1/pkg
5015	350 μL Receiver Plate Spacer Block	1/pkg
5016	Replacement Accessory Kit (Includes O-Ring, gasket, and allen wrench)	1/pkg