Section 8
The Vacuum Regulator
The vacuum regulator allows complete control of the negative pressures inside
the Aurum vacuum manifold, allowing optimum levels of negative pressure to
be achieved 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 the vacuum
filter flask, the regulator measures the difference between ambient pressure
and the pressure within the vacuum manifold in inches of mercury (“ Hg). The
regulator is capable of measuring negative pressures up to -30” Hg (Figure 7).
For other purification kits, please consult the appropriate protocols for
recommended negative pressures.
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” Hg. 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 (Figure 3).
11
10
Fig. 6. Column purification protocol overview.
Aurum Vacuum Manifold: Cat. #732-6470
For more information, call Technical Service
at 1-800-4BIORAD (1-800-424-6723). Visit us
on the Web at
www.bio-rad.com/aurum/
Neutralized
lysate
Vacuum
manifold
Vacuum
filter flask
Vacuum
regulator
Vacuum
source
Elution
solution
Fig. 7. Vacuum regulator with clear representation
of the gauge.
Aurum
™
Vacuum
Manifold: Column
Purification
Protocol Overview
For complete protocol, consult instruction manual.
Tubing provided is 4 ft long and must be cut into
appropriate pieces before proceeding.
Vacuum Setup Conditions
Vacuum source to manifold
1.
Use tubing to connect vacuum source to
right side
of vacuum regulator unit.
2.
Use tubing to connect
left side
of vacuum
regulator to filter flask.
Rubber stopper of vacuum filter flask must
contain a hole to accommodate serological
(or comparable) pipet.
Vacuum trap must contain additional sidearm
for connection to regulator.
3.
Use tubing to connect filter flask to nozzle
of manifold top.
Vacuum Purification Conditions
When ready to purify
4.
Place A stage in center of manifold base.
5.
Place waste collection tray on top of A stage.
6.
Place manifold top on base to ensure
complete contact.
7.
Place the column adapter plate (CAP) on
manifold top.
Ensure luer is end up.
Ensure unused luer fittings are tightly closed.
8.
Place desired Aurum vacuum column(s) into
open luer ends.
9.
Proceed with appropriate purification protocol
for column processing.
Spin Elution Conditions
When ready to elute
10.
Proceed with appropriate spin elution step as
recommended by protocol.
* The Aurum vacuum manifold and stages should
be rinsed with water after each use.
Aurum Vacuum Manifold: Cat. #732-6470
For more information, call Technical Service
at 1-800-4BIORAD (1-800-424-6723). Visit us
on the Web at
www.bio-rad.com/aurum/
Bulletin 2671 US/EG Rev A 01-229 0501
Vacuum
manifold
Vacuum
filter flask
Vacuum
regulator
Vacuum
source
