Aurum
™
Vacuum Manifold
Instruction Manual
For technical service, call your local Bio-Rad office, or
in the US, call 1-800-4BIORAD (1-800-424-6723)
Bio-Rad Laboratories
2000 Alfred Nobel Dr.
Hercules, CA USA 94547
(510) 741-1000
1-800-424-6723
4110115 Rev A
Table of Contents
Section 1 Introduction . . . . . . . . . . . . . . . . . . . . . . . . . .1
Section 2 Kit Components . . . . . . . . . . . . . . . . . . . . . . . .1
Section 3 Care and Maintenance . . . . . . . . . . . . . . . . . . .2
Section 4 The Aurum Vacuum Manifold . . . . . . . . . . . . . .3
Section 5 Instrument Setup and Use for 96-well Plates . . . . .5
Section 6 The Column Adaptor Plate (CAP) . . . . . . . . . . .7
Section 7 Instrument Setup and Use for the
Column Adaptor Plate . . . . . . . . . . . . . . . . . . .8
Section 8 The Vacuum Regulator . . . . . . . . . . . . . . . . . .11
Section 9 Troubleshooting Guide . . . . . . . . . . . . . . . . . .13
Section 10 Ordering Information . . . . . . . . . . . . . . . . . . .15
Section 1
Introduction
The Aurum vacuum manifold and column adaptor plate (CAP), comprise a
patent-pending 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:
Vacuum manifold consisting of:
- manifold top
- manifold base
Column adaptor plate with luer caps
Vacuum regulator
A stage
B stage
Waste collection tray
Replacement luer caps (4)
Tubing (5/16” ID)
1
Section 3
Care and Maintenance
The Aurum vacuum manifold and CAP should be stored clean and dry at
room temperature. After each use rinse the instruments with distilled
water, and wipe dry with paper towels or air dry. Never use alcohols 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 may
promote uneven contact between manifold parts and the purification
plate or CAP. Check the tubular and flat silicone gaskets for wear before
and after use of the manifold. Should the upper flat gaskets become
worn out, replacements (Cat. 732-6471) can be ordered by contacting
Bio-Rad Customer Service at 1-800-4BIORAD (1-800-424-6723). In the
event that the bottom tubular gasket requires replacement, a length of
1/8" OD silicone tubing may be used.
2
Section 4
The Aurum Vacuum Manifold
The Aurum vacuum manifold is designed to accommodate the 96-well plates
used in Bio-Rad's kits, such as the Aurum plasmid 96 kit. The manifold may
also be compatible with other manufacturers' kits. For column processing, the
column adaptor plate is required. The vacuum regulator is included and
strongly recommended for measuring negative pressure, as well as for
gradually increasing negative pressure at different periods throughout the
various 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 occasionally be required to
facilitate seating. In the event that the tubular silicone gasket becomes
detached, it can be easily reinserted into the track on the underside of the
manifold top.
Each Aurum vacuum manifold is supplied with two stages of different heights:
a short A stage and tall B stage. Both stages are used with the etched letters
A and B facing up. On the top side of the A stage is a shallow depression, in
which the waste collection tray is placed. During sample loading and washing,
the A stage centers the waste collection tray underneath the Aurum
purification plate or the CAP. The taller B stage centers and elevates a 96-well
sample collection plate during elution from the purification plate. A groove
outlining the perimeter of the top side of the B stage accommodates the
bottom skirt of the collection plate.
3
Fig. 1. Aurum vacuum manifold kit components.
Manifold top
Manifold base Vacuum regulator
A stage
Replacement luer caps
Column adaptor plate
Tubing (5/16” ID)
B stage Waste collection tray
4
Section 5
Instrument Setup and Use for
96-well Plates
Tubing provided is 4 ft. long and must be cut in appropriate pieces before
proceeding.
Vacuum setup conditions
1. Cut tubing into 3 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. Finally, use the last piece of tubing to connect the filter flask to the nozzle
of the manifold.
Note: The Aurum vacuum regulator is strongly recommended to enable
full control of the negative pressure within the manifold.
Vacuum purification conditions
6. Remove the manifold top and place the A stage in the depression in the
center of the manifold base. The etched letter A should be facing up.
7. Place the waste collection tray on top of the A stage. Replace the
manifold top on the base, ensuring complete and uniform contact
between the manifold top and base.
8. Place the desired 96-well plate on the manifold top, and follow the
instructions in the protocol for the appropriate purification kit.
Vacuum elution conditions
9. When eluting from the purification plate, remove the A stage and waste
collection tray from the Aurum vacuum manifold. Place the B stage on the
manifold base, with the etched letter B facing up. Place a clean 96-well
microtiter collection plate securely on top of the B stage and replace the
manifold top.
10. After finishing the elution, rinse the Aurum vacuum manifold with water,
and air dry or wipe with paper towels.
5
Fig. 2. Instrument setup and use for plate and column
processing. 2a: 96-well purification, 2b: 96-well elution, 2c: CAP
purification
Fig. 3. Vacuum Setup Conditions
a.
b. c.
6 7
Section 6
The Column Adaptor Plate (CAP)
The Column Adaptor Plate (CAP) interfaces with the Aurum vacuum manifold
to convert the manifold from a plate- to a column-processing system (Figure 6).
The CAP has 18 black luer fittings in a 6 x 3 array and comes supplied with
additional caps. Up to 18 Aurum miniprep columns, 18 Aurum midiprep
columns, or 9 Aurum maxiprep 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.
* If more than 6 maxi columns are used, empty the waste collection tray
before the wash step.
Fig. 4. 96-well purification protocol overview.
Bulletin 2667 US/EG Rev A 01-185 0501
Fig. 5. Column Adaptor Plate with and without Aurum vacuum
manifold. 5a: Blow-up view of luer connection between the Bio-Rad mini and
column adaptor plate, 5b: CAP with individual columns (maxi’s, midi’s & mini’s).
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/
Vacuum
manifold
Vacuum
filter flask
Vacuum
regulator
Vacuum
source
Aurum
™
Vacuum
Manifold: 96-Well
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 desired Aurum 96-well plate on manifold
top and proceed with appropriate protocol.
Vacuum Elution Conditions
When ready to elute
8. Remove A stage and waste collection tray
from manifold.
9. Place B stage in center of manifold base.
10. Place clean 96-well collection plate on top
of B stage.
11. Replace manifold top on base to ensure
complete contact.
12. Proceed with appropriate elution step as
recommended by protocol.
* The Aurum vacuum manifold and stages should
be rinsed with water after each use.
a.
b.
Section 7
Instrument Setup and Use for the
Column Adaptor Plate
Preparing the Aurum manifold:
Tubing provided is 4 ft. long and must be cut in appropriate pieces
before proceeding.
Vacuum set-up conditions (please see Figure 3, page 4).
1. Cut tubing into 3 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. Finally, use the last piece of tubing to connect the filter flask to the
nozzle of the manifold.
Note: The Aurum vacuum regulator is strongly recommended to
enable full control of the negative pressure within the manifold.
Vacuum purification conditions
6. Insert the CAP (luer ends up) into the depression in the vacuum
manifold top. Ensure that the CAP rests evenly on the gasket.
7. 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.
8. Insert the luer ends of the desired Aurum columns into the available
luer fittings, ensuring a tight fit. The manifold is now ready for
column processing according to the vacuum protocol of the
appropriate column purification kit.
9. After finishing the purification, rinse the CAP and Aurum vacuum
manifold with distilled water, and air-dry or wipe with paper towels.
Vacuum elution conditions
10. When ready to elute, proceed with the appropriate spin elution step as
recommended by the protocol.
11. After finishing the elution, rinse the Aurum vacuum manifold with water,
and air dry or wipe with paper towels.
98
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
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
Use A stage for
waste collection
Purification plate or CAP Press down gently on
not fully seated 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
13
Table 1. Pressure unit conversions
To convert from inches of mercury (
““
Hg) to Multiply by:
millimeters of mercury or torr (mm Hg, torr) 25.4
millibar (mbar) 33.85
atmospheres (atm) 0.03342
pounds per square inch (psi) 0.4912
kilopascals (kPa) 3.385
Table 2. Vacuum manifolds and their preparative formats
12
Vacuum Purification Mini Midi Maxi 96
Aurum Vacuum Manifold • • • •
Promega Vac-Man •
Promega Vac-Man 96 •
QIAGEN QIAvac 96 •
QIAGEN QIAvac 24 •
Millipore vacuum manifold •
14 15
Section 10
Ordering Information
Catalog # Description
732-6470
AAuurruumm vvaaccuuuumm mmaanniiffoolldd
, includes column adaptor plate,
4 replacement luer caps, A and B stages, waste collection
tray, vacuum regulator and gauge, tubing, protocol overview
and instruction manual.
732-6400
AAuurruumm ppllaassmmiidd mmiinnii kkiitt
, 100 preps, includes 100 plasmid
binding columns, 100 capless collection tubes (2.0 ml),
100 capped sample tubes (1.5 ml), reagents, protocol
overview and instruction manual.
732-6440
AAuurruumm ppllaassmmiidd mmiiddii kkiitt
, 20 preps, includes 20 lysate
filtration columns, 20 plasmid binding columns, 20 adaptor
caps, reagents, protocol overview and instruction manual.
732-6450
AAuurruumm ppllaassmmiidd mmaaxxii kkiitt
, 10 preps, includes 10 lysate
filtration columns, 10 plasmid binding columns, 10 adaptor
caps, reagents, protocol overview and instruction manual.
732-6460
AAuurruumm ppllaassmmiidd 9966 kkiitt
, 2 x 96 well preps, includes 2 grow
blocks, 2 grow membranes, 2 binary units (2 lysate filtration
plates and 2 plasmid binding plates), 2 collection plates,
reagents, protocol overview and instruction manual.
Problem Possible Cause Possible Solution
Worn manifold gasket Replace gasket
Incorrect negative Vacuum source connected Connect vacuum
pressure from vacuum to left side of vacuum to right side of vacuum
regulator
Clogging of purification Insufficient negative See Problem “Insuffi-
plate or CAP columns pressure in vacuum cient negative pressure
manifold in vacuum manifold”
above
Purification problem Refer to troubleshooting
seated properly section of appropriate
kit
Plasmid prep performs Proteinaceous debris in See Problem Low
poorly in enzymatic prep (if A
260/280
< 1.7) A
260/280
ratio
reactions
Plasmid is degraded Use endA
-
strain (e.g.
JM109, DH5α, XL1-
Blue)
Ethanol contamination in Add 1-3 minutes to the
prep (eluate volumes purge time after the
> 55 ml) wash step
Remove all traces of
Wash
Solution from the drip
directors of the plasmid
binding plate by blotting
on paper towels prior to
elution
/
