Bio-Rad Vacuum Manifold User Manual

Aurum

™

Vacuum Manifold

Instruction Manual

For technical support, call your local Bio-Rad office, or
in the US, call 1-800-4BIORAD (1-800-424-6723).

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Table of Contents

Section 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . .1

Section 2 Kit Components . . . . . . . . . . . . . . . . . . . . . . . . .1

Section 3 Care and Maintenance . . . . . . . . . . . . . . . . . . . .2

Section 4 Aurum™ Vacuum Manifold . . . . . . . . . . . . . . . . .3

Section 5 Vacuum Regulator . . . . . . . . . . . . . . . . . . . . . . .4

Section 6 Vacuum Manifold Setup . . . . . . . . . . . . . . . . . . .5

Section 7 Use for 96-Well Plates . . . . . . . . . . . . . . . . . . . . . . . .6

Section 8 Use for the Column Adaptor Plate (CAP) . . . . . .7

Section 9 Troubleshooting Guide . . . . . . . . . . . . . . . . . . . .8

Section 10 Ordering Information . . . . . . . . . . . . . . . . . . . .10

Section 1
Introduction

The Aurum™ vacuum manifold and column adaptor plate (CAP)` comprise a
system which allows a single vacuum manifold to be used with either 96-well
plates or columns with luer ends. Separate manifolds are no longer required
for different preparative formats; the Aurum vacuum manifold and CAP ensure
fast, high-quality sample preparation in both formats, while maintaining the
simplicity of handling associated with vacuum processing. The uniquely
designed vacuum regulator allows for complete control of negative pressure
within the Aurum vacuum manifold.

Section 2
Kit Components

The Aurum™ vacuum manifold contains the following components
as shown in Figure 1:

• Vacuum manifold consisting of:

- manifold top

- manifold base

• Column adaptor plate

• Vacuum regulator

• Stage

• Luer caps

• Tubing (

5

/

16

” ID)

1

Section 3
Care and Maintenance

The Aurum™ vacuum manifold should be stored clean and dry at room
temperature. After each use rinse with distilled water and wipe dry with
paper towels or air dry. Never use alcohol or other organic reagents to
clean the manifold. Never use abrasive materials to clean the manifold.

Gasket surfaces should be carefully rinsed to remove salts, which could
inhibit contact between manifold parts and the column adaptor plate
(CAP). Check the tubular and flat silicone gaskets for wear before and
after use of the manifold. In the event that the bottom tubular gasket
requires replacement, a length of

1

/

8

" OD silicone tubing may be used.

2

Section 4
Aurum™ Vacuum Manifold

The Aurum vacuum manifold is designed to accommodate the 96-well plates
used in Bio-Rad's Aurum total RNA 96 kit. The manifold may also be
compatible with other manufacturers' kits. For column processing, the column
adaptor plate (CAP) is required.

The CAP allows the Aurum vacuum manifold to to be used for processing
columns with a luer fitting. The CAP has 18 black luer fittings in a 6 x 3 array.
Up to 18 Aurum miniprep columns can be accommodated on the CAP
without the need for connectors or other manifold accessories. The CAP will
also accommodate other columns with luer ends.

When vacuum is applied to the manifold, the CAP should self-seat, forming an
airtight seal without the need to press down on the CAP. However, the
application of gentle downward force may occasionally be required to facilitate
seating.

The vacuum regulator is included and strongly recommended for measuring
negative pressure, and for gradually increasing negative pressure during
different protocols.

The Aurum manifold consists of two primary parts: the manifold base and top.
The top has an attached tubular silicone gasket, allowing the manifold base
and top to seal when a vacuum is introduced. Attached to the opening in the
manifold top is a flat silicone gasket, enabling Aurum 96-well purification plates
or the CAP to establish an airtight seal with the manifold top when a vacuum
is applied. Collectively these two gaskets facilitate self-seating of all
components, reducing the need to apply external pressure to the manifold to
initiate vacuum. However, under certain conditions application of gentle
downward force on the plates or CAP may be required to facilitate seating. In
the event that the tubular silicone gasket becomes detached, it can easily be
reinserted into the track on the underside of the manifold top.

Each Aurum vacuum manifold is supplied with a stage that centers and
elevates a 96-well sample collection plate during elution from the purification
plate. The stage should be placed with the printed "this side up" in the base
with the groove side down to allow for airflow.

3

Fig. 1. Aurum vacuum manifold kit components.

Manifold top

Manifold base with

quick connect fitting

Vacuum regulator

A stage

Luer caps

Column adaptor plate (CAP)

Tubing (

5

/

16

” ID)

Stage Waste collection tray

Section 5
Vacuum Regulator

The vacuum regulator (see Figure 2) allows complete control of the negative
pressures inside the Aurum™ vacuum manifold, allowing optimum levels of
negative pressure for efficient sample preparation. This is particularly important
when a gradual increase in negative pressure is required during 96-well plate
purifications. Attached in-line between the vacuum source and filter flask, the
regulator measures the difference between ambient pressure and pressure
within the vacuum manifold in inches of mercury (inHg). The regulator is
capable of measuring negative pressures up to –30 inHg. See Table 1 for
pressure unit conversions.

At various points in the protocols, it is necessary to release the vacuum inside
the manifold so that plates and columns can be removed. To release the
negative pressure, rotate the regulator dial fully counter-clockwise until the
gauge reads < 1 inHg. The vacuum is restored inside the manifold by rotating
the control dial clockwise until the desired negative pressure is attained.

The regulator control dial is located off-center, toward the right side of the
regulator. The vacuum source is attached to the right hose fitting and the
vacuum manifold is attached to the left fitting (see Figure 3).

Table 1. Pressure unit conversions

To convert from inches of mercury (inHg) to:

Multiply by:
millimeters of mercury or torr (mmHg, torr) 25.4
millibar (mbar) 33.85
atmospheres (atm) 0.03342
pounds per square inch (psi) 0.4912
kilopascals (kPa) 3.385

Fig. 2. Vacuum regulator and gauge.

4

Section 6
Vacuum Manifold Setup

Tubing provided is 4 ft long and must be cut in appropriate pieces before
proceeding.

Prior to setup, you may ensure that the gauge pointer is adjusted to zero by
removing the lens cover, followed by turning the adjustment pin located
beneath the dial face.

Vacuum setup

1. Cut tubing into three pieces of appropriate length.

2. Use one piece of tubing to connect the vacuum source to the right side
of the vacuum regulator.

3. Use another piece of tubing to connect the left side of the vacuum
regulator to the sidearm of the filter flask.

4. Place a rubber stopper with hole into the mouth of the filter flask. Insert
a serological pipette (or comparable) into the hole of the stopper.

5. Snap in the black sealed end of the quick connect fitting into the manifold
base.

6. Finally, use the last piece of tubing to connect the filter flask to the quick
connect fitting of the nozzle of the manifold.

Fig. 3. Vacuum setup conditions.

5

6 7

Section 8
Use for the Column Adaptor Plate
(CAP)

Manifold wash setup (see Figure 5)

1. Insert the CAP (luer ends up) into the depression in the vacuum manifold
top. Ensure that the CAP rests evenly on the gasket.

2. Insert the luer ends of the desired columns into the available luer fittings,
ensuring a tight fit.

3. Close the unused luer fittings with the caps provided. Close caps by
rotating clockwise until light resistance is encountered. Excessive
tightening of a cap may cause the luer fitting to dislodge when the cap is
removed.

4. The manifold is now ready for column processing according to the
vacuum protocol of the appropriate column purification kit.

5. When ready to elute, proceed with the appropriate spin elution step as
recommended by the protocol.

6. After finishing the elution, rinse the CAP and Aurum vacuum manifold
with water, and air dry or wipe with paper towels.

Fig. 5. Setup and use for column processing.

Section 7
Use for 96-Well Plates

Manifold wash setup (see Figure 4A)

1. Place the manifold top on the base, ensuring complete and uniform
contact between the manifold top and base.

2. Place the desired 96-well purification plate on the manifold top and apply
the recommended vacuum pressure for your application.

Manifold elution setup (see Figure 4B)

3. When eluting from the purification plate, place the stage on the manifold
base, with the etched "this side up" facing up. Place a clean 96-well
microtiter collection plate securely on top of the stage and replace the
manifold top.

Note: If desired, waste can be collected into an optional pipet tip box lid
instead of draining directly into the vacuum trap.

4. After use, rinse the Aurum™ vacuum manifold with water and air dry or
wipe with paper towels. Failure to rinse the vacuum manifold at the end of
each use will cause the manifold to become cloudy and pitted.

Fig. 4. Setup and use for 96-well plate processing.

A) 96-well wash; B) 96-well elution.

A.

B.

Section 9
Troubleshooting Guide

Problem Possible Cause Possible Solution

Insufficient negative Open vacuum regulator Close vacuum regulator
pressure in vacuum by turning completely
manifold clockwise

Manifold top not seated Ensure uniform contact
properly on base between manifold top

and base

Purification plate or CAP Press down gently on
not fully seated the corners of the plate

or CAP to seal

Unused CAP luer fittings Close unused luer
not closed completely fittings completely with

caps

Gasket residue preventing Rinse manifold with
even contact between distilled water
manifold parts

Lower gasket misaligned Press gasket back into

groove

Physical damage to Replace vacuum
manifold or CAP (e.g., manifold or CAP
chips and cracks)

Purge step in purification If closure of wells or
protocol columns (e.g., with

sealing tape) restores
negative pressure, no
corrective action
needed

Excessive noise Purification plate or CAP Press down gently on
during vacuuming not fully seated plate or CAP to seat

Gasket residue preventing Rinse manifold with
even contact between distilled water
manifold parts

Worn manifold gasket Replace gasket

Problem Possible Cause Possible Solution

Incorrect negative Vacuum source Connect vacuum
pressure from connected to left to right side of vacuum
vacuum regulator side of vacuum

Clogging of Insufficient negative See “Insufficient
purification plate pressure in vacuum negative pressure
or CAP columns manifold in vacuum manifold”

problem above

98

Section 10
Ordering Information

Catalog # Description

732-6470 Aurum Vacuum Manifold

732-6820 Aurum Total RNA Mini Kit

732-6830 Aurum Total RNA Fatty and Fibrous Tissue Kit

732-6800 Aurum Total RNA 96 Kit

732-6400 Aurum Plasmid Mini Kit

10