PureLink Vacuum Manifold

User Manual

Corporate HeadquartersInvitrogen Corporation1600 Faraday AvenueCarlsbad, CA 92008T: 1 760 603 7200F: 1 760 602 6500E: [email protected]

For country-specific contact information visit our web site at www.invitrogen.com

PureLink™ Vacuum Manifold For rapid, parallel processing of nucleic acids Catalog no. K2110-01

Version A 24 April 2007 25-1014

ii

Table of Contents Kit Contents and Storage .............................................................................v Additional Products ....................................................................................vi

Introduction .............................................................................................1 Overview........................................................................................................ 1

Methods....................................................................................................4 General Guidelines ....................................................................................... 4 Vacuum Manifold - General Assembly and Use...................................... 8 Vacuum Manifold Assembly Using Filter and Binding Plates............. 10 Vacuum Manifold Assembly Using Binding Plates Only..................... 12

Appendix ................................................................................................14 Troubleshooting.......................................................................................... 14 Technical Support and Product Warranty .............................................. 16

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iv

Kit Contents and Storage

Shipping and Storage

The PureLink™ Vacuum Manifold is shipped at room temperature.

Upon receipt, and after each use, store the manifold clean, and dry at room temperature.

System Components

The components included with the PureLink™ Vacuum Manifold are listed below.

Component Quantity

PureLink™ Vacuum Manifold Base with valve

1 each

PureLink™ Vacuum Manifold Lid 1 each

Waste Tray 2 each

Spacer Sets • MTP/Multi-96 well plates

• Round-well blocks • Square-well blocks • Microtube racks

1 set 1 set 1 set 1 set

v

Additional Products

Additional Products

The following additional products are available from Invitrogen for use with the PureLink™ Vacuum Manifold. A variety of nucleic acid purification kits may be used with the manifold.

For more details on these products, visit our website at www.invitrogen.com, or contact Technical Support (see page 16).

Product Quantity Catalog no.

PureLink™ Vacuum Regulator 1 each K2110-02

PureLink™ Quick96 Plasmid Kit 4 x 96 preps 24 x 96 preps

K2110-04 K2110-24

PureLink™ 96 Genomic DNA Kit 4 x 96 preps K1821-04

PureLink™ 96 Total RNA Purification Kit 384 preps 12173-011

PureLink™ 96 HQ Mini Plasmid DNA Purification Kit

4 x 96 preps K2100-96

ChargeSwitch® PCR Clean-Up Kit 100 preps 960 preps

CS12000 CS12000-10

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Introduction

Overview

Introduction The PureLink™ Vacuum Manifold is designed for rapid manual purification of nucleic acids. The manifold reduces sample handling to a minimum by allowing direct parallel and simultaneous processing of up to 96 samples using collection plates, deep-well blocks, or microtubes. The use of vacuum eliminates the need for time-consuming pipetting and centrifugation. Note: The PureLink™ Vacuum Manifold is not designed for use with automated liquid handling systems.

Manifold Features

The PureLink™ Vacuum Manifold offers the following features and advantages:

• Allows rapid and simultaneous processing of up to 96 samples of nucleic acids

• Compatible with a variety of nucleic acid purification kits (see below)

• Provides spacer sets, enabling the processing and collection of samples with different types of plates, blocks, or tubes (see page 2)

• Eliminates cross-contamination of samples when properly assembled

• Resistant to routine laboratory chemicals (see page 3)

Manifold Applications

The PureLink™ Vacuum Manifold is suitable for use with the following nucleic acid purification kits:

Application Invitrogen Kit

Plasmid Purification PureLink™ Quick96 Plasmid Kit PureLink™ 96 HQ Mini Plasmid Kit

RNA Purification PureLink™ 96 Total RNA Purification Kit

Genomic DNA Purification

PureLink™ 96 Genomic DNA Kit

PCR Clean-Up ChargeSwitch® PCR Clean-Up Kit

Continued on next page

1

Overview, Continued

Description of Parts

A description of the PureLink™ Vacuum Manifold parts is provided below.

Manifold Lid: The manifold lid is placed on top of the

manifold base. Plates are inserted into center section of lid. Lid dimensions: L = 17.2 cm, W= 15.3 cm, H = 1.8 cm

Manifold Lid

Manifold Base: The manifold base includes the gasket and the valve. The base holds the spacers sets, waste trays and purification plates. Base dimensions: L = 16.7 cm, W = 14.8 cm, H = 7.2 cm • Valve: The valve attaches to the vacuum line for

pressure reduction. Refer to the diagram below to open and close the valve.

• Gasket: The gasket compresses with the application of vacuum to seal the manifold lid to the manifold base.

“Open” Valve “Closed” Valve

Gasket

Manifold BaseValve

Waste Trays: The waste trays are placed inside the manifold base to collect liquid waste produced during purification.

Waste Trays Spacer Sets: Spacer sets are used with microtiter plates (MTP)/96-well plates, round-well blocks, square-well blocks, and microtube racks. The type of spacer set used will depend on kit specifications. The spacers slide into the grooves in the manifold base for correct spacing and alignment of plates.

Spacer Sets


2

Overview, Continued

Chemical Properties of the Manifold

The table below presents the materials and resistance properties of the PureLink™ Vacuum Manifold. Note: Although the manifold parts are resistant to certain chemicals, we recommend avoiding prolonged exposure to these chemicals. Do not expose the manifold parts to the chemicals listed in the not resistant column.

Manifold Part Part

Material Parts are resistant to: Parts are not

resistant to:

Manifold Base Spacer Sets Waste Containers

Polyacrylic

Manifold Lid Polyvinyl Chloride (PVC)

Manifold Valve Stainless steel, other metals

Dilute Acetic Acid Ethanol Guanidine-HCl Sodium Chloride (NaCl) Dilute Sodium Hydroxide (NaOH) Sodium Dodecyl - Sulfate (SDS)

Acetone Phenol Toluene Chlorine bleach Any apolar organic solvents

3

Methods

General Guidelines

Introduction General guidelines for using the PureLink™ Vacuum Manifold are described below. Review this section before using the manifold.

Guidelines for assembling the vacuum manifold for the following applications are described:

General assembly Page 8

Plasmid purification using Page 10 filter and binding plates

Nucleic acid purification using Page 12 binding plates only

• The vacuum manifold operates under pressure. For your protection, wear a laboratory coat, gloves, and safety glasses when operating the manifold.

• The manifold has a maximum vacuum reduction threshold of -800 millibars (mbars) (800 mbars below atmospheric pressure). For safety reasons, do not exceed -800 mbars. Exceeding -800 mbars of vacuum pressure will void the product warranty.

• Use only the recommended vacuum pressure stated in your purification kit manual. Using higher than the recommended vacuum pressure may cause sample splattering or inefficient DNA binding, while using a lower than recommended vacuum pressure affects the elution, resulting in lower recovery.


4

General Guidelines, Continued

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Follow the recommendations below to obtain the best results when using the PureLink™ Vacuum Manifold.

• Always operate the manifold on a secure, flat lab bench or work area.

• Ensure proper use and maintenance of the manifold (see page 7).

• Calibrate the vacuum pressure prior to using the manifold (see page 9).

• Use a vacuum regulator during high-throughput processing to attain consistent purification results and achieve specific pressure ranges.

• Use the correct spacer set for your purification needs (see page 2). Insert the spacers completely into the grooves of the manifold base to prevent cross-contamination and allow for proper alignment and spacing of the top and bottom plates.

Vacuum Sources

The PureLink™ Vacuum Manifold can be used with the following vacuum sources:

• House Vacuum (ducted to the laboratory)

• Vacuum Pump

• Water Aspirator

Pressure Conversions

The table below provides information to convert pressure from millibars into other commonly used units.

If converting from millibars to… Then multiply by…

atmospheres (atm) 0.000987

bars 0.001

inches of Mercury (in. Hg) 0.0394

millimeters of Mercury (mm Hg) 0.75

pounds per square inch (psi) 0.0145


5


PureLink™ Vacuum Regulator

The PureLink™ Vacuum Regulator (see page vi) measures the pressure difference between the inside of the vacuum manifold and the outside (atmospheric pressure). Units on the PureLink™ Vacuum Regulator are in mbars and psi. Note: Pressure conversions are provided on page 5.

To set up and use the PureLink™ Vacuum Regulator with the PureLink™ Vacuum Manifold:

1. Ensure the vacuum source is turned off and the valve on the manifold base is closed (see valve description on page 2).

2. Connect the regulator between the manifold base and the vacuum source:

• Connect one piece of vacuum tubing from the valve on the manifold base to one of the valves on the vacuum regulator.

• Connect a second piece of vacuum tubing from the second regulator valve to the valve on the vacuum source.

3. Turn on the vacuum source and use the regulator knob to adjust the vacuum pressure.

• To increase the vacuum pressure, turn the regulator knob counter-clockwise.

• To decrease or turn off the vacuum pressure, turn the regulator knob clockwise.

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6

Regulator Knob

Regulator Valves

Store the vacuum regulator at room temperature.

Dry the valve after each use to prevent permanent damage to the regulator caused by condensation and rust build-up.



Care and Maintenance of the Vacuum Manifold

Care and maintenance guidelines for the PureLink™ Vacuum manifold are provided below.

Maintaining the Manifold

• Wash and disassemble the parts of the manifold after each use (see below). Do not autoclave the manifold or its components.

• Store the manifold clean and dry, at room temperature.

• Keep the manifold valve clean and dry to ensure optimal vacuum reduction.

• Do not exceed -800 mbars of vacuum reduction.

• Do not apply silicone or vacuum grease to the gasket or any other part of the vacuum manifold. The gasket on the manifold base compresses to seal the lid to the base when the correct amount of vacuum is applied. If the gasket is damaged, see Troubleshooting on page 14.

• Avoid exposing any part of the manifold to harsh chemicals (see page 3).

• Do not drop the vacuum manifold.

Washing the manifold

1. After each use, disassemble the manifold parts. Remove the spacer set from the base immediately after each use. Salts and buffers can dry behind or underneath a removable part and may prevent the future use or removal of the part, or may damage the manifold.

2. Thoroughly wash all manifold parts after each use to remove salts and buffers. Do not use solvents, abrasives or chlorine bleach when washing the manifold. Note: It is important to clean the gasket on the manifold base after each use; however, we recommend that you do not remove the gasket from the manifold base, unless it is damaged.

3. Always dry the manifold parts completely with an absorbent towel. To prevent rusting of the metal components, do not let the manifold air dry.

7

Vacuum Manifold - General Assembly and Use

Introduction Instructions for the general assembly and use of the PureLink™ Vacuum Manifold are provided below.

General Assembly

Follow the steps below to assemble the PureLink™ Vacuum Manifold.

Place Manifold Lidonto Manifold Base.

Insert Spacer Setand Waste Tray intoManifold Base.

AssembledVacuum Manifold.

Manifold Lid

Spacer Set

Manifold Base

Waste Tray

Valve

1. Place the manifold on a flat, secure lab bench or work area.

2. Place a waste tray inside the manifold base and insert the appropriate spacers (notch side up) completely inside the grooves of the manifold base (see page 2).

3. Place the manifold lid on top of the manifold base.

4. Connect the manifold to a suitable vacuum source (see page 5) and calibrate the vacuum pressure (see page 9). Ensure the manifold valve is closed before turning on the vacuum source (see page 2 for valve use).

Ventilating the Manifold

Ventilating the manifold is required at certain steps during purification. Ventilating also allows for easy removal of purification plates or blocks, and the manifold lid.

To ventilate the manifold, close the manifold valve and wait for 10-20 seconds. Note: During purification using PureLink™ plates, ventilate the manifold completely after each vacuum step for 60-90 seconds.


8

Vacuum Manifold - General Assembly and Use, Continued

Calibrating Vacuum for Use with 96-Well Plates

To attain optimal purification results, calibrate the vacuum pressure prior to using the PureLink™ Vacuum Manifold.

A vacuum regulator is required for this step. Note: For 96-well plates, we recommend using a vacuum pressure of -200 to -400 mbars (-6 to -12 inches Hg or -150 to -300 mm Hg).

To calibrate the vacuum pressure:

1. Assemble the vacuum manifold (described on page 8), and place an unused 96-well plate in the manifold lid. Seal the plate with Foil Tape.

2. Ensure the valve on the manifold base is closed (see page 2).

3. Connect a vacuum regulator between the manifold base and the vacuum source (described on page 6).

4. Turn on the vacuum source and adjust the pressure with the vacuum regulator.

5. Open the valve on the manifold base after attaining the desired pressure.

6. Once all steps are complete, close the manifold valve and ventilate the manifold to release the vacuum seal and prepare for the purification steps (see page 8 for ventilation).

The vacuum manifold is now ready for use.

The vacuum pressure may need to equilibrate and may rise slightly above your preset value. Allow the vacuum to run for a few minutes before beginning the purification.

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Invitrogen is not responsible for any injury or damage caused by the use of this unit when operated for purposes which it is not intended. Use of the manifold in a manner not specified in this manual, will void the warranty offered on this unit. All repairs and service to this unit should be performed by Invitrogen.

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Vacuum Manifold Assembly Using Filter and Binding Plates

Introduction Assembling the PureLink™ Vacuum Manifold for a plasmid

kit with 96-well filter and binding plates, e.g., PureLink™ Quick96 Plasmid Kit, is performed in two steps.

The first step filters the lysate and binds the DNA.

The second step washes the samples and elutes the DNA. For details on how to perform each step, refer to the purification kit manual.

Assembly with Filter and Binding Plates

Instructions for assembling the vacuum manifold using filter and binding plates are described below.


Place Binding Plateinto Base. PlaceManifold Lid on top.Insert Filter Plateinto Lid.

Assembled VacuumManifold with FilterPlate in lid andBinding Plate in base.

Filter Plate

Manifold Base

Waste Tray

Spacer Set

Valve

Filter Plate

Manifold Lid

Binding Plate

Discard Filter Plate.Remove ManifoldLid and Binding Plate.Place Binding Plateinto Manifold Lid.

BindingPlate

1. Connect the manifold to a vacuum source (page 5). Close the manifold valve.

2. Insert a waste tray and the appropriate spacer set (notch side up) into the manifold base (see page 2).

3. Place the binding plate onto the notches of the spacers in the manifold base. Place the manifold lid on top and insert the filter plate into the lid.

4. Transfer lysates into the filter plate. Align the plates and apply vacuum (open the manifold valve) to filter the lysate onto the binding plate.

5. Ventilate the manifold to release the plates (see page 8).

6. Discard the filter plate. Remove the manifold lid and the binding plate from the manifold base.

7. Replace the lid and insert the binding plate into the lid.

8. Apply vacuum to bind the DNA to the binding plate.


10

Vacuum Manifold Assembly Using Filter and Binding Plates, Continued

Assembly with Wash and Elution Plates

Assembly instructions using wash plates* and elution plates are described below.

* Not all purification kits include a wash plate. Refer to the purification kit manual for detailed instructions.

Place Wash Plate orElution Plate intoManifold Base. InsertBinding Plate intoManifold Lid.

Assembled VacuumManifold withBinding Plate in Lidand Wash or ElutionPlate in Base.

Assembled VacuumManifold withBinding Plate only.

ManifoldLid

Binding Plate

ElutionPlate

WashPlate

Binding Plate 1. From the previous step, remove the manifold lid and the inserted binding plate from the base. Place the wash plate* onto the notches of the spacers in the manifold base.

2. Replace the manifold lid with the inserted binding plate onto the manifold base. Align the binding plate and the wash plate.

3. Perform the necessary wash steps and apply vacuum.

4. Ventilate the manifold to release the plates.

5. Remove the manifold lid and binding plate. Remove and discard the wash plate (if applicable).

6. Place the elution plate onto the notches of the spacers in the manifold base.

7. Replace the manifold lid with the inserted binding plate on top of the manifold base. Align the binding plate and the elution plate.

8. Perform the elution step and apply vacuum.


10. Disassemble and clean the manifold (see page 7).

11

Vacuum Manifold Assembly Using Binding Plates Only

Introduction Instructions for assembling the PureLink™ Vacuum

Manifold using a nucleic acid purification kit with 96-well binding plates only, e.g., PureLink™ Genomic DNA Kits, are described for binding and washing steps, and elution steps. For details on how to perform each step, refer to the purification kit manual.

Assembly with Binding and Wash Plates

Assembling the manifold using binding and wash plates* is described below. * Not all purification kits include a wash plate. Refer to the purification kit manual for detailed instructions.

Assembled VacuumManifold withBinding Plate in lidand Wash Plate inBase.

Place Wash Plate*into Manifold Base.Insert Binding Plateinto Manifold Lid.


ManifoldLid

Binding Plate

Wash Plate

Manifold Base

Waste Tray

Spacer Set

Valve

1. Connect the manifold to a vacuum source (page 5). Close the manifold valve.

2. Insert a waste tray and the appropriate spacers (notch side up) into the manifold base.

3. Place the manifold lid on top of the manifold base and insert the binding plate into the lid.

4. Transfer lysates to the binding plate and apply vacuum (open the manifold valve) to bind the DNA to the binding plate.

5. Ventilate the manifold to release the plates (see page 8).

6. Remove the lid and binding plate and place the wash plate* onto the notches of the spacers in the manifold base.

7. Replace the manifold lid with the inserted binding plate onto the base. Align the binding plate and the wash plate.

8. Perform the necessary wash steps and apply vacuum.



12

Vacuum Manifold Assembly Using Binding Plates Only, Continued

Assembly with Binding and Elution Plates

Assembling the manifold using binding and elution plates is described below.

Place Elution Plateinto Manifold Base.Replace Manifold Lidwith Binding Plate.

Assembled VacuumManifold withBinding Plate andElution Plate.

Remove Manifold Lidand Binding Plate.Discard Wash Plate.*

ElutionPlate

ManifoldLid

Binding Plate

ManifoldLid

Binding Plate

Wash Plate

1. From the previous binding step, remove the manifold lid and the binding plate from the manifold base. Remove and discard the wash plate* (if applicable).

2. Place the elution plate onto the notches of the spacers in the manifold base.

3. Replace the manifold lid with the inserted binding plate on top of the manifold base. Align the binding plate and the elution plate.

4. Perform the elution step and apply vacuum.


6. Disassemble and clean the manifold (see page 7).

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Appendix

Troubleshooting

Introduction The table below describes solutions to possible problems you may experience with the vacuum manifold. For additional assistance, please contact Invitrogen Technical Support (see page 16).

Problem Cause Solution

Gasket • Determine if the gasket is working properly by inserting a microtiter plate (MTP) with a closed, rectangular bottom into the manifold lid. The microtiter plate must cover the gasket of the vacuum manifold completely. Apply vacuum to the manifold and test the vacuum. The MTP should be fixed now by vacuum. You may need to apply some gentle pressure to the MTP with your hands.

• Check to see that the gasket is clean of salts and buffers, and is not damaged.

• If the gasket is damaged, contact Technical Support for a replacement (see page 16).

Vacuum manifold is damaged

• Check for cracks on the surface of and around the manifold base, valve, and lid.

Inadequate vacuum build-up or vacuum pressure leak

Vacuum pressure is not adequate

• Ensure the valves on the manifold base and vacuum source are clear of debris or salt build-up.

• Ensure that the vacuum tubing is not clogged or damaged.

• Change to a new vacuum source if the current one is too weak.


14

Troubleshooting, Continued

Problem Cause Solution Incorrect plate used

• Ensure the plate you are using fits inside the vacuum manifold. Note: Using an incorrect plate prevents a tight seal between the lid and the base.

Inadequate vacuum build-up or vacuum pressure leak, continued Incorrect spacer

set used • Use only the spacer set designed for

the type of plate used for sample collection (see page 2).

• Make sure the spacers are completely inserted inside the manifold base.

Plates are not properly aligned

• Align the top plate in the lid completely with the bottom plate in the base. Once vacuum pressure is applied, the channels of the top plate will move into the wells of the bottom plate, preventing cross-contamination.

Sample leaking and cross-contamination

Manifold lid is not tightly sealed

• With the vacuum pressure on, apply a few seconds of pressure to the manifold lid or top plate with your hands to compress the gasket.

• Make sure the gasket is clean and is not damaged (see previous page).

• Use plates that are the correct size for the vacuum manifold.

• Use the appropriate spacer set for the type of plate you are using (see page 2 for spacer information).

• Confirm you have adequate vacuum pressure (see previous page).

15

Technical Support and Product Warranty

Web Resources

Visit the Invitrogen website at www.invitrogen.com for:

• Technical resources including manuals, vector maps and sequences, application notes, MSDSs, etc.

• Complete technical support contact information. • Access to the Invitrogen Online Catalog. • Additional product information and special offers

Contact Us For more information or technical assistance, call, write, fax, or

email. Additional international offices are listed on our website (www.invitrogen.com).

Corporate Headquarters Invitrogen Corporation 1600 Faraday Avenue Carlsbad, CA 92008 USA Tel: 1 760 603 7200 Tel (Toll Free): 1 800 955 6288 Fax: 1 760 602 6500 E-mail: [email protected]

Japanese HeadquartersInvitrogen Japan LOOP-X Bldg. 6F 3-9-15, Kaigan Minato-ku, Tokyo 108-0022 Tel: 81 3 5730 6509 Fax: 81 3 5730 6519 E-mail: [email protected]

European Headquarters Invitrogen Ltd Inchinnan Business Park 3 Fountain Drive Paisley PA4 9RF, UK Tel: +44 (0) 141 814 6100 Tech Fax: +44 (0) 141 814 6117 E-mail: [email protected]

Certificate of Analysis

Certificate of Analysis (CofA) for the product is available on our website at www.invitrogen.com/cofa.

PureLink™ Vacuum Manifold Warranty

Invitrogen warrants that the PureLink™ Vacuum Manifold will be free from defects in material and workmanship for a period of one year from date of purchase. If a defect is present, Invitrogen will, at its option, repair, replace, or refund the purchase price of this product at no charge to you, provided it is returned during the warranty period. This warranty does not apply if the product has been damaged by accident, abuse, misuse or misapplication, or from ordinary wear and tear. This warranty shall be limited to the replacement of defective products. It is expressly agreed that this warranty will be in lieu of all warranties of fitness and in lieu of the warranty of merchantability.

©2007 Invitrogen Corporation. All rights reserved. For research use only. Not intended for any animal or human therapeutic or diagnostic use.

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User Manual

Corporate HeadquartersInvitrogen Corporation1600 Faraday AvenueCarlsbad, CA 92008T: 1 760 603 7200F: 1 760 602 6500E: [email protected]

For country-specific contact information visit our web site at www.invitrogen.com

PureLink Vacuum Manifold

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