Hybridizer, HybriLinker and Multidizer Hybridization Ovens Operating Instructions HB-1000 Hybridizer HM-4000 Multidizer HL-2000 HybriLinker IMPORTANT: To familiarize yourself with the correct operation procedures, read these instructions before operating your UVP Hybridizer, Multidizer or HybriLinker system. _____________________________________________________________________________________ UVP, LLC 2066 W.
Hybridization Ovens Table of Contents Table of Contents ........................................................................................... 2 Introduction ..................................................................................................... 3 Hybridization Oven Specifications .................................................................. 4 Setup Instructions ........................................................................................... 6 Operating Instructions .....
Hybridization Ovens Introduction The HB-1000 Hybridizer is a cost-effective, high-performance, 10-bottle hybridization oven with state-of-the-art microprocessor temperature and variable speed controls. The HL-2000 HybriLinker™ System is designed with a combination of hybridizer and crosslinker components. This equipment is intended to be used in the laboratory environment. The HM-4000 Multidizer™ System is designed as a multifunction unit with a variety of motion options.
Hybridization Ovens Hybridization Oven Specifications HB-1000 Hybridizer Hybridization Oven Part Numbers 95-0030-01 95-0030-02 95-0030-03 Volts/Hz 115V/60Hz 230V/50Hz 100V/50Hz Specifications Net Weight: Temperature: Temperature Stability: Temperature Uniformity: Temperature Display: Heating Element: Rotation Speed: Bottle Capacity: Dimensions: 43 lbs Ambient +10°C to 99.9°C Inside Oven: +/-0.3°C to 68°C Inside Bottles: +/-0.1°C to 68°C Inside Oven: +/-0.1°C to 68°C Inside Bottles: +/-0.
Hybridization Ovens HM-4000 Multidizer Hybridization Oven Part Numbers 95-0340-01 95-0340-02 95-0340-03 Volts/Hz 115V/60Hz 230V/50Hz 100V/50Hz Specifications Net Weight: Temperature Display: Dimensions: Upper Chamber Temperature: Temperature Stability: Temperature Uniformity: Heating Element: Shaker Tray Speed: Carousel Rotation: 75 lbs (34 kg) LCD Exterior 17.5W x 28.5H x 15D in. (445 x 724 x 381 mm) Ambient +10°C to 80°C Inside Oven: +/-0.3°C to 68°C Inside Bottles: +/-0.
Hybridization Ovens Setup Instructions Two different bottle sizes are available for use within the hybridizer. The rotary wheel carousel can hold up to twenty (20) 35 x 150 mm bottles or ten (10) 35 x 300 mm bottles. In addition, combinations of both bottle sizes may be used at the same time. • To insert bottle holder carousel (Fig. 1), slide the two wheels onto the rod and tighten the thumb screws (Fig. 2). Insert the rod into the holder on the right wall of the hybridizer chamber.
Hybridization Ovens Installing the Rollers and Optional Trays To insert rollers in the hybridization oven carousel, slide the rollers into the receptacle on the left side of the chamber first, followed by the right side. To install the Reciprocating Shaker Tray (standard with the HM-4000), place the tray over the rollers in the hybridization oven. To install the Rocker Tray (optional on all units), remove the metal carousel from the chamber.
Hybridization Ovens Operating Instructions Please note that this equipment is not intended for interconnection with any other devices. Use of this equipment other than intended may create a safety hazard and/or malfunction. Using the Hybridization Oven 1. Place the unit on a level working surface and provide adequate room in front of the door to open it easily. 2. Plug the female end of the power cord into the unit. 3.
Hybridization Ovens Calibrating the Temperature Reading The hybridizer is calibrated at the UVP factory. UVP recommends temperature recalibration be performed at the UVP factory as recalibration by the user may void the warranty. Obtain an RGA (Returned Goods Authorization) number from UVP prior to returning a unit to the factory. Call UVP’s customer service department in the USA in Upland, California at (800) 452-6788 or (909) 946-3197 or in the UK in Cambridge, UK at +44(0)1223420022.
Hybridization Ovens connectors, ensure that the proper configuration of the male connector or the plug has been properly connected to the power cord. 4. Turn the On/Off switch to the On position. NOTE: WHEN TURNED ON THE CROSSLINKER DEFAULTS TO THE LAST USED UV EXPOSURE SETTING. 5. 6. The last UV exposure setting will now be displayed on the LCD and the last function setting will be noted by glowing red spot(s) on the display panel. Place your sample requiring exposure into the chamber. 7.
Hybridization Ovens User-Set UV Time Exposure Setting your own UV time exposure can be achieved as follows: 1. 2. 3. Press “TIME” on the tactile touchpad and then set your requirements by pressing the corresponding numbers. Your time exposure settings will be displayed flashing on the LCD. Remember that your time exposure settings are displayed in minutes and tenths of a minute. If your settings are correct, press “ENTER” on the tactile touchpad. Press “START”.
Hybridization Ovens call UVP. 11. This calibration process should be repeated following any unsuccessful recalibration. If calibration is unsuccessful, replace the new UV wavelength tubes with the previously-removed old wavelength tubes. 12. Successful recalibration to the NEW UV wavelength changes the values in the microprocessor and the numeric setting value is displayed on the LCD. To return to original or another wavelength, the proper UV calibration sensor is needed.
Hybridization Ovens Care and Cleaning NOTE: Always unplug the unit from the electrical supply before cleaning or drying the unit. The units are built to provide you with trouble-free operation. To ensure correct operation: • Wipe ANY water from inside and outside the unit with a soft cloth or sponge. • Use soap and water with a soft cloth or sponge to clean the unit. • Do not allow chemicals to remain on the unit’s surfaces. • Never clean the unit with abrasive pads or cleaners.
Hybridization Ovens Replacement Parts Replacement parts lists are provided below for the HB-1000 Hybridizer and HL-2000 HybriLinker systems. Repairs or replacement other than as specified in the following procedures should only be completed by authorized service personnel. Replacement Parts for HB-1000 Hybridizer DESCRIPTION Borosilicate Glass Bottle, with Cap and O-Ring Large, 35 x 300mm Cap and O-Ring (for all Bottle sizes) Bottle Holder Carousel Carousel Drive Shaft Rod Drip Tray QTY. NEEDED PART NO.
Hybridization Ovens Replacement Parts for HM-4000 Multidizer DESCRIPTION Borosilicate Glass Bottle, with Cap and O-Ring Large, 35 x 300mm Cap and O-Ring (for all Bottle sizes) Bottle Holder Carousel Carousel Drive Shaft Rod Drip Tray (for Lower Chamber) QTY. NEEDED PART NO.
Hybridization Ovens Accessories Bottles: Borosilicate glass bottles with polypropylene cap and PTFE seal: Bottle, Small, 35 x 150 mm 07-0194-01 Bottle, Oversize Call UVP Bottle Carousels: Carousel Kit for Four Oversized Bottles 76-0066-02 Carousel for (10) 50ml Conical Bottles 76-0075-01 Carousel, Acrylic (for HM-4000 upper chamber) 76-0089-01 Capacity: (4) 35 x 300mm, (8) 35 x 150mm, or (8) 50mm Conical Other Equipment: Rocker Tray (for all models), 12.5W x 10D in.
Hybridization Ovens Maintenance, Repair and Technical Assistance UVP offers technical support for all of its products. If you have any questions about the product’s use, operation or repair, please call or fax UVP Customer Service at the following numbers: UVP, LLC In the US, toll-free (800) 452-6788 or (909) 946-3197 Fax: (909) 946-3597 2066 W. 11th Street, Upland, CA 91786 USA Ultra-Violet Products Ltd. In Europe/UK, call +44(0)1223-420022 Fax: +44(0)1223-420561 Ultra-Violet Products Ltd.
Hybridization Ovens Hybridization Techniques There are really two main steps to a hybridization reaction: hybridizing two strands of complementary DNA, and detection of the hybridized DNA. Nucleic acid hybridization is a mechanism where strands of DNA in a single stranded state have their complements bind together. The proximity of the DNA strands to each other determines the frequency of the binding events and, in fact, successful binding is proportional to their concentration.
Hybridization Ovens 3. Salt Concentration and Temperature Nucleic acid requires salt (monovalent cations) to reduce the ionic effects of the phosphate backbone, and heat as a form of non-denaturing kinetic energy. Because the salt concentration and temperature effect each other, knowing the thermostability of the hybrid probe is helpful. Hybridization rate varies directly with the sodium ion concentration between 0.03 and 1.2 M. Most protocols run between 0.5 and 1.1 M Sodium.
Hybridization Ovens Protocol 1: Random Priming Method for Tagging DNA with Fluorescein-Labeled Nucleotide and Others This method uses DNA polymerase to incorporate Fluorescene-11- dUTP into double stranded DNA probes. This protocol can be used to incorporate any tagged nucleotides. Equipment • Micropipettes and tips • Boiling water bath • 1.5 mL Microcentrifuge tubes • Microcentrifuge • Cap lock for Microcentrifuge tube • Water bath set to 37°C Reagents • Deionized, sterile water • EDTA, 0.
Hybridization Ovens Protocol 2: Hybridization to Nylon or Nitrocellulose Hybridization to nylon or nitrocellulose membranes containing Nucleic Acid is accomplished by adding single-stranded probe to the membranes that have been previously incubated with prehybridization solution. The prehybridization and hybridization solutions both contain buffers designed to prevent adventitious binding of the probe to the filters. Reagents and Equipment Prehybridization/hybridization solution [45% formamide, 5X SSPE (0.
Hybridization Ovens Radioactive Probes Additionally you will need: • • • X-Ray film holder X-Ray film Intensifying screen Procedure 1. Remove blot from hybridization tube and transfer to Tupperware container 2. Rinse briefly in 50 °C 0.1X SSC, 0.1% SDS. 3. Remove this solution to radioactive waste and wash 4. Wash blot three more times in the same solution. 5. After the final wash, dry blot on filter paper for 10 minutes.
Hybridization Ovens Protocol 3: Chemiluminescence Detection: HRP-Tagged, Alkaline Phosphatase (AP) Probes or Antibody Conjugates Equipment • Clear plastic cling-wrap or Clear transparent sheet protector • UVP EC3 or AC1 Darkroom with Cooled CCD camera • Pipette Reagents for Chemiluminescence • ECL™ (or other) detection reagent 1 • ECL™ (or other if required) detection reagent 2 • Membrane following hybridization. Procedure 1. Mix equal volumes of detection reagents 1 and 2 2.
