Bio-Rad 785 Vacuum Blotter User Manual

Model 785

Vacuum Blotter

Instruction

Manual

For Technical Support

Call Your Local Bio-Rad Office or

in the U.S. Call 1-800-4BIORAD

(1-800-424-6723)

Table of Contents

Section 1 General Information ................................................................................................1

1.1 Safety/Definition of Symbols ....................................................................................1

1.2 Introduction to Model 785 Vacuum Blotter................................................................1

1.3 List of Parts ................................................................................................................1

1.4 Features ......................................................................................................................1

1.5 Specifications..............................................................................................................2

Section 2 Description of Major Parts ......................................................................................3

2.1 Base Unit ....................................................................................................................3

2.2 Vacuum Stage ............................................................................................................3

2.3 Porous Vacuum Plate..................................................................................................3

2.4 Window Gasket ..........................................................................................................3

2.5 Sealing Frame ............................................................................................................4

2.6 Vacuum Regulator ......................................................................................................4

2.7 Vacuum Pump ............................................................................................................4

2.8 Lid ..............................................................................................................................4

Section 3 Operating Instructions ............................................................................................5

3.1 Vacuum Blotter Setup ................................................................................................5

3.2 Preparation for Vacuum Transfer................................................................................5

3.3 Gel Pretreatment ........................................................................................................5

3.4 Vacuum Transfer ........................................................................................................7

3.5 Post Transfer ..............................................................................................................8

Section 4 Cleaning and Maintenance ......................................................................................8

4.1 Cleaning......................................................................................................................8

4.2 Replacement Parts ......................................................................................................8

Section 5 Troubleshooting Guide ............................................................................................9

5.1 Operational Troubleshooting ......................................................................................9

5.2 Southern Hybridization Troubleshooting ................................................................10

5.3 Repairing Cracked Gels ............................................................................................11

Section 6 Equipment and Accessories ..................................................................................12

6.1 Model 785 Vacuum Blotter and Accessories............................................................12

6.2 Electrophoresis Reagents..........................................................................................12

6.3 Blotting Reagents......................................................................................................13

Section 7 Appendix..................................................................................................................13

7.1 Facts About Vacuum Transfer ..................................................................................13

7.2 Required Solutions and Equipment ..........................................................................15

Section 1 General Information

1.1 Safety/Definition of Symbols

Caution

This instrument is intended for laboratory use only.

This product conforms to the “Class A” standards for electromagnetic emissions intended for laboratory equipment applications. It is possible that emissions form this product may interfere with some sensitive appliances when placed nearby or in the some circuit as those applicances. The user should be aware of this potential and take appropriate measures to avoid interference.

1.2 Introduction to the Model 785 Vacuum Blotter

The Model 785 Vacuum Blotter is a nucleic acid transfer apparatus which rapidly transfers DNA or RNA from agarose gels onto nylon membranes by using low vacuum. The Model 785 Vacuum Blotter simplifies every step of the blotting process. In addition, the Model 785 Vacuum Blotter is constructed for long term heavy use. The following protocols have been thoroughly tested and have the continuing technical support of Bio-Rad Laboratories.

1.3 List of Parts

Each Model 785 Vacuum Blotter unit comes with the items listed below. Check to make sure that all components are present. Also note any damage to the instrument that may have occurred during shipping. Inform Bio-Rad Laboratories if any items are missing or damaged.

Item Quantity

Base Unit 1 Vacuum Stage 1 Porous Vacuum Plate 1 Window Gasket – precut 15 x 15 cm 2 – precut 15 x 20 cm 2 – blank 2 Sealing Frame 1 Lid 1

Offered Accessories

Vacuum Regulator 1 Vacuum Station 1

1.4 General Description

The Vacuum Blotter has several innovative features which simplify the nucleic acid blotting process and facilitate complete transfer. The Model 785 Vacuum Blotter features include:

• Easy-to-use spring latches which make assembly and disassembly quick. No screws or nuts to tighten.

• Vacuum Stage and Porous Vacuum Plate create an even vacuum pressure throughout the

transfer area, providing even transfers and highly reproducible results.

• Large Vacuum Stage format permits large or small gel transfer using the blank Window

Gasket or standard gel transfer using the precut Window Gasket.

• Green Window Gasket allows you to easily see the gel/gasket overlap for easy alignment.

• Vacuum Regulator facilitates accurate vacuum pressure using Bio-Rad’s Vacuum Pump or almost any vacuum source.

• Durable molded construction for long term heavy use.

• Base Unit and Sealing Frame designed for easy alignment of gel and membrane.

1.5 Specifications

Dimensions

Overall outer size 21” (L) x 12” (W) x 5” (H) Gel capacity (maximum size) 20 cm x 25 cm

Unit weights

Complete assembly 8.5 lb Lid weight 1.5 lb Sealing frame assembly 1.5 lb Porous vacuum support 0.25 lb

Materials

Base Mindel-S plastic Springs Stainless steel Posts Delrin plastic Vacuum stage Mindel-S plastic Window gasket Flexible green vinyl Porous vacuum plate Polypropylene Sealing frame Glass-filled Mindel-S O-ring Silicone rubber Lid Acrylic Tubing Tygon (1/2” OD x 1/4” ID)

Vacuum regulator pump

Dimensions

Overall outer size 16” (L) x 9” (W) x 8” (H)

Weights

With pump 9.5 lb Without pump 3.9 lb

Materials

Valve Polyvinylchloride Fittings Nylon Housing Acrylic Interior pump components PTFE

Ratings

Vacuum pump 20 inches Hg (max)/250 Torr Regulator 0-25 inches Hg (max)/125 Torr

Electrical

Input voltage range 100–120 VAC/50-60 Hz/1.5 A

Fuses

Internal Thermal Fuse

220–240 VAC/50 Hz/0.75 A

Environmental

Operating 50 °F (10 °C) to 90 °F (32 °C) temperature

Storage 32 °F (0 °C) to 140 °F (60 °C) temperature

Cautions

1. Not designed to handle liquids or heavy vapors. If pump does get liquid inside, open valve to max position and let run for 15 minutes to allow interior components to dry.

2. If valve knob loosens up during operation, wrap threads with PTFE tape to increase thread engagement.

3. Pump will become hot after a few hours of continuous use. This is normal and should not be a cause for concern.

Section 2 Description of Major Parts

2.1 Base Unit

The Base Unit is the foundation of the Model 785 Vacuum Blotter. The plastic used to make the Base Unit is an off-white color with a slight texture. The Base Unit is made of a durable and chemically resistant mindel-S plastic. In low concentrations, acids and bases 0.25 N HCl or 0.5 N NaOH will not damage the Base Unit or the Vacuum Stage. The Base Unit supports the Vacuum Stage on springs and catches the transfer solution during disassembly.

30–80% humidity

10–90% humidity

2.2 Vacuum Stage

The Vacuum Stage is supported by springs on the Base Unit. The spring action on the Vacuum Stage is used to lock and unlock the Sealing Frame. The center portion of the Vacuum Stage has a grooved grid pattern. This grid pattern helps distribute the vacuum pressure evenly across the Porous Vacuum Plate. In the center of the grid pattern is a port. This port is the vacuum inlet and is connected to vacuum tubing underneath the Vacuum Stage. Surrounding the grid area is a soft silicon Reservoir Seal O-ring that provides a water-tight seal to the upper buffer chamber.

2.3 Porous Vacuum Plate

The Porous Vacuum Plate is a white polypropylene sheet. The Porous Vacuum Plate is used to support the transfer membrane and the gel. Along with the grid pattern on the Vacuum Stage, the Porous Vacuum Plate helps distribute the vacuum pressure evenly.

2.4 Window Gasket

The Precut Window Gasket is a green vinyl sheet with a window cut out of the center. Refer to Section 3.2 for instructions on how to cut the Blank Window Gasket. The Window Gasket is used to mask off the remaining Porous Vacuum Plate not covered by the gel. The Window Gasket forces the vacuum through the gel/membrane complex by making a watertight seal where the gel overlaps the Window Gasket. The Window Gasket also seals in the vacuum along the Reservoir Seal O-ring when the Sealing Frame is compressed onto the Vacuum Stage.

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