Page finder Important Information ..........ii Waste Electrical and Electronic Equipment (WEEE) ......viii Introduction ............1 Specifications............3 Set-up instructions ..........4 Operating instructions ..........6 Care and maintenance .........10 Customer service information ........10 Hybridization techniques ........11 Experimental protocols .........13 References ............20 Ordering information ..........21 •...
Important Information – English • Pozor! Protože tento nástroj může vyvinout dostatečný napětí a proud, který má vyrábět a • If this equipment is used in a manner not speci- smrtiacej šok, péče musí být vykonávána v jeho fied by Hoefer, Inc. the protection provided by the provoz. equipment may be impaired. • Tento nástroj je určen v souladu s EN61010-1:2001 • This instrument is designed for indoor laboratory elektrické bezpečnostní normy. Přesto, že by měly use only. být použity pouze řádně vyškolený operátorů. Číst • Only accessories and parts approved or supplied celé toto ruční před použitím nástroje a použití by Hoefer, Inc. may be used for operating, main- pouze v souladu s pokyny. taining, and servicing this product. • Přístroj musí být vždy používají se na výkonu zemi • Warning! Because this instrument can develop vést šňůra správně zemněny k zemi na síti výústce. sufficient voltage and current to produce a lethal • Využití pouze nepoškozené elektrické dráty a shock, care must be exercised in its operation. vybavení pro napětí budete používat. Všechna • This instrument is designed in accordance with zařízení spojené s vysokým napětím by měla být v the EN61010-1:2001 electrical safety standard.
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• Om toe te staan voldoende afkoelen, verzeker equilibrate til omgivende temperatur ved tageen dat de luchtopeningen van het instrument niet instrumentets fra et koldere til et varmere miljø. bedekt zijn. • At tillade tilstrækkelig afkøling, forsikrer, at Tärkeää Tietoa – Finnish lufthullerne af instrumentet er ikke dækket. • Jos tätä varusteita käytetään tavassa ei määritetty Belangrijke Informatie – Dutch Hoeferille, Inc. suojelu ehkäisty varusteille saattaa olla avuton. • Indien deze uitrusting in een manier wordt gebruikt die niet door Hoefer is gespecificeerd, Nv. • Tämä väline suunnitellaan sisälaboratoriokäytölle de bescherming die door de uitrusting is verzorgd vain. kan worden geschaad. • Vain lisävarusteet ja osat hyväksyivät tai toimitti • Dit instrument is voor binnenlaboratoriumgebruik Hoeferin oheen, Inc.:ää voi käyttää käyttämiselle, enkel ontworpen. valvoalle, ja servicing tämä tuote. • Enkel onderdelen en delen keurden goed of • Varoittaminen! Koska tämä väline voi kehittää riit- leverden door Hoefer, Nv. kan voor het bedienen tävä jännitteen ja virran tuottaa kuolettavan järky-...
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Information Importante – French • Dieses Instrument wird für den Innenlaborge- brauch nur dafür entworfen. • Si cet équipement est utilisé dans une manière pas • Nur Zusätze und Teile genehmigten oder lieferten spécifié par Hoefer, Inc. la protection fourni par durch Hoefer, Inc kann für das Funktionieren, das l’équipement pourrait être diminuée. Aufrechterhalten, und die Wartung dieses Produk- • Cet instrument est conçu pour l’usage de labora- tes verwendet werden. toire intérieur seulement. • Die Warnung! Weil dieses Instrument genügend • Seulement les accessoires et les parties ont Stromspannung und Strom entwickeln kann, um approuvé ou ont fourni par Hoefer, Inc. pourrait einen tödlichen Stoß zu erzeugen, muss Sorge in être utilisé pour fonctionner, maintenir, et entrete- seiner Operation ausgeübt werden. nir ce produit. • Dieses Instrument wird in Übereinstimmung mit • Avertissant! Parce que cet instrument peut dével- dem EN61010-1:2001 elektrischen Sicherheitsstan- opper la tension et le courant suffisants pour dard dafür entworfen. Dennoch sollte es nur von produire un choc mortel, le soin doit être exercé richtig erzogenen Maschinenbedienern verwendet dans son opération. werden. Lesen Sie dieses komplette Handbuch vor • Cet instrument est conformément conçu à...
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• Dette instrumentet er utformet i samsvar med usato per operare, per mantenere, e per revisionare EN61010-1:2001 elektrisk sikkerhetsstandard. questo prodotto. Likevel burde bli brukt det bare av skikkelig utdannede operatører. Les denne hele håndboken • Avvertendo! Perché questo strumento può før bruking instrumentet og bruken bare gi til sviluppare il voltaggio sufficiente e la corrente di instruksjonene. produrre una scossa letale, la cura deve essere esercitata nella sua operazione. Questo strumento • Instrumentet må alltid bli brukt med jorden blyet è disegnato conformemente all’EN61010-1:2001 av kraftkabelen som riktig ha blitt jordet til jord på la norma di sicurezza elettrica. Tuttavia, dovrebbe hovedledningen utløp. essere usato degli operatori solo correttamente • Bruker bare uskadd elektrisk ledningsfremføring addestrati. Leggere questo manuale intero prima og utstyr som er spesifikk for spenningene du vil di usare lo strumento e l’uso solo secondo le bruke. All utstyr koplet til høyspenning burde være istruzioni. i samsvar med EN61010-1:2001. • Lo strumento deve essere sempre usato col • Beholder instrumentet som tørker og rengjør som piombo di terra della spina di alimentazione corret- mulig. Visk regulært med et mykt, fuktig stoff. La tamente hanno messo a terra alla terra alla presa di instrumentet tørker komplett før bruk. corrente principale. • Driver instrumentet i ekstrem fuktighet ikke (oven- • Usa il filo metallico e l’apparecchiatura solo intatti...
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• Permitir esfriar suficiente, assegura que as abertu- ras do instrumento não são cobertas. • Nie prowadzą do instrumentu w skrajnych wilgotności (powyżej 80%). Zapobiec kondensacji Información Importante – Spanish najmu przez jednostkę równoważyć do temper- atury pokojowej przy podejmowaniu instrumentu • Si este equipo es utilizado en una manera no espe- z chłodniejsze w cieplejszych środowiska. cificado por Hoefer, S.a. la protección proporcio- • Aby umożliwić wystarczające chłodzenia, nado por el equipo puede ser dañada. zapewniają, że rozcięcia of the instrument nie • Este instrumento es diseñado para el uso interior objęte ubezpieczeniem. del laboratorio sólo. Sólo accesorios y partes aprobaron o suministraron por Hoefer, S.a. puede Informações Importantes – ser utilizado para operar, para mantener, y para Portuguese atender a este producto. • Advirtiendo! Porque este instrumento puede • Se este equipamento é usado numa maneira não desarrollar voltaje y corriente suficientes para especificada por Hoefer, Inc. que a protecção forne-...
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• Permitir refrigeración suficiente, asegure que las aberturas del Viktig Information – Swedish • om denna utrustning används i ett sätt som inte har specificeras av Hoefer, Inc. skyddet tillhan- dahöll vid utrustningen kan skadas. • Detta instrument formges för inomhuslaborato- rium användning bara. • Bara medhjälpare och delar godkände eller lever- erade vid Hoefer, Inc. kan användas för fungera, underhålla, och servicing denna produkt. • varna! Därför att detta instrument kan utveckla tillräcklig spänning och ström att producera en dödlig stöt, måste övas omsorg i dess funktion. • Detta instrument formges i överensstämmelse med EN61010-1:2001 elektriska säkerheten stan- darden. Icke desto mindre, bör det användas bara av riktigt utbildade operatörer. Läs denna hela handbok före använda instrumentet och använd bara enligt undervisningarna. • Instrumentet måste alltid användas med jorden blyet av kraften repet riktigt grounded till jorden på det huvudutloppet. • Använder bara undamaged elektrisk tråd och utrustning specifik för spänningarna du ska använda. All utrustning kopplats som till hög spän- ning skulle vara i överensstämmelse med EN61010- 1:2001.
Waste Electrical and Electronic Equipment (WEEE) English This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately. Please contact an authorized representative of the manufacturer for information concerning the decommissioning of your equipment.
IMPORTANT! Please read these instructions before The HB1000 Hybridization Oven can be set to operating your Hybridization operate anywhere from 10 to 18 RPM, enabling Oven to familiarize yourself with consistent saturation of samples, whether it be operation procedures.
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WARNING! There may be build up of pressure within the hybrid- ization bottles when they are taken from ambient to hybridization temperature. To help in relieving some of this pressure, preheat the solutions and bottles. Also, be sure bottles are opened at the same temperature at which the hybridization was done.
Set-up instructions Two different bottle sizes are available for use in Note: Always use an even number of bottles and load the hybridization oven. The bottle holder carousel bottles at opposite points to can hold up to twenty (20) 35 × 150 mm bottles balance the wheels of the bottle or ten (10) 35 ×...
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Installing the optional rocker tray coupling sleeve Remove the bottle holder carousel from the chamber. Position the rocker tray in the chamber so that the coupling arm is located on the right side. Place the coupling sleeve over the motor drive shaft so that the pins on the inside of the coupling Fig 4.
Operating instructions Using the Hybridization Oven Place the unit on a level work surface and provide adequate room in front of the door for easy access, as well as the back for air ventilation. Plug the female end of the power cord into the back of the oven.
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Setting the temperature controls Whenever power is applied to the controller, the software revision number is displayed for several seconds. While the software revision number is being displayed, the intensity of the display digits alternates between full and half brightness, and on some models the two outside decimals points blink on and off.
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Calibrating the temperature reading The hybridizer is calibrated at the factory. Hoefer, Inc. recommends temperature recalibra- tion be performed at the factory as recalibration by the user may void the warranty. Obtain a RA (Return Authorization) number from Hoefer, IMPORTANT! Request a copy Inc.
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Bottle care Hybridization bottles are made of lead-free borosilicate glass, which provides excellent long- term reliability. Each bottle includes a polypro- pylene cap and Teflon seal. ® • It is important to check your bottles regularly for chips, stress fractures and cracks. If these occur, the bottle must be discarded.
• Never clean unit with acetone or chloroform. Customer service information Technical service and repair Hoefer, Inc. offers complete technical support IMPORTANT! Request a copy of for all of our products. If you have any ques- the Hoefer, Inc. “Health and Safety Declaration”...
Hybridization techniques There are two main steps to a hybridization reaction: 1) hybridizing two strands of DNA carrying complementary sequences, and 2) detec- tion of the hybridized DNA. Proximity of the DNA strands determines the frequency of the binding event and, therefore, success is depen- dent on the DNA concentrations.
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Mechanisms of nucleic hybridization Hybridization occurs with a process called nucleation whereby the two separate nucleic acid strands come into close proximity of each other. A duplex region is formed where a minimum of three bases of one strand comple- ments to those on the second strand.
Experimental protocols There are many different protocols available on the web, in journals, and in textbooks. We refer- ence several at the end of this section. 1. Concentration of species Target: The ideal quantity of target molecule depends on the complexity of the target nucleic acid. The amplified DNA, for example, may have been expressed in large or small amounts.
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2. Length of probe The goal is to increase hybridization efficiency while minimizing background. The larger the probe, the more efficiently it binds target. The smaller the probe, however, an ever greater number of probe molecules is present per unit amount (ng). In most cases probes range from 20 –...
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4. Aqueous or denaturing hybridization buffer If hybridization takes place in an aqueous salt environment of 0.8 to 1.2 M salt, the T (the m½ temperature at which half of the duplex molecules will dissociate under a given set of conditions) can be 90 °C.
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Reagents • Deionized, sterile water • EDTA, 0.5 M • DNA polymerase, Klenow fragment, 4 – 5 units/μL • Nucleotide mix (300 μM each of dATP, dCTP, dGTP and 60 μM dTTP) • Random nonamer (9-mer) primers, 2.5 μg/μL in water •...
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100 mg/mL of denatured salmon sperm DNA]. Mix SDS will precipitate at room well and remove aggregates before use. temperature but remain in • Hoefer hybridization bottle(s) and caps solution at 37 °C. • 15 mL plastic tube • Boiling water bath •...
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AVOID pouring the probe directly onto the blot. 6. Incubate in the Hoefer HB1000 Hybridization Oven for 6 to 8 hours at 42 to 56 °C. Washing the blot •...
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Non radioactive probes 1. Wash blots in 2X SSC, 0.1% SDS for 3 minutes at room temperature (repeat one). 2. Wash filter in 0.15X SSC, 0.1% SDS for 15 minutes at 50 °C (repeat once). 3. Store blots in 2X SSC at room temperature. Radioactive probes 1.
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