BIO RAD Aurum User manual

Brand: BIO RAD

Model: Aurum

Type: User manual

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)

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.

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.

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

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.

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

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.

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.

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).

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

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

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

BIO RAD Aurum User manual

Brand: BIO RAD

Model: Aurum

Type: User manual

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BIO RAD Aurum User manual

BIO RAD Aurum User manual

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