HE-PLUS User Manual – English
Contents
1. Usa il filo metallico e apparecchiatura solo intatti elettrici specifici per i voltaggi che lei usera Tutta Vapparecchiatura collegata all alto voltaggio dovrebbe essere conformemente a EN61010 1 2001 Tiene lo strumento come secco e pulito come possibile Pulire regolarmente con un morbido per spegnere il panno Lasciare lo strumento asciuga completamente prima dell uso O piv Non opera lo strumento nell umidita estrema al di sopra di 80 Evitare la condensazione lasciando l unit equilibra alla temperatura ambiente quando portare lo strumento da un piu freddo a un ambiente piu caldo Di permettere raffreddare sufficiente assicura che gli sbocchi dello strumento non sono coperti Viktig Informasjon Norwegian Hvis dette utstyret blir brukt i en m te ikke spesifisert ved Hoefer Inc beskyttelsen som ha blitt git av utsty ret kan bli svekket Dette instrumentet er utformet for innendors labora toriumbruk bare Bare tilbeh r og deler godkjente eller forsynte ved Hoefer Inc kan bli brukt for drive vedlikeholde og betjene dette produktet Varsler Fordi dette instrumentet kan utvikle tilstrek kelig spenning og strom til produsere et dodelig sjokk m bli vd bekymring i dets drift Dette instrumentet er utformet i samsvar med EN61010 1 2001 elektrisk sikkerhetsstandard Likevel burde bli brukt det bare av skikkelig utdannede oper atorer Les denne hele h ndboken for bruking instru2. German Wenn diese Ausr stung gewisserma en nicht angege ben durch Hoefer Inc verwendet wird kann der durch die Ausr stung zur Verf gung gestellte Schutz verschlechtert werden Dieses Instrument wird f r den Innenlaborgebrauch nur daf r entworfen Nur Zus tze und Teile genehmigten oder lieferten durch Hoefer Inc kann f r das Funktionieren das Aufrechterhalten und die Wartung dieses Produktes verwendet werden Die Warnung Weil dieses Instrument gen gend Strom spannung und Strom entwickeln kann um einen t dli chen Sto zu erzeugen muss Sorge in seiner Operation ausge bt werden Dieses Instrument wird in bereinstimmung mit dem EN61010 1 2001 elektrischen Sicherheitsstandard daf r entworfen Dennoch sollte es nur von richtig erzo genen Maschinenbedienern verwendet werden Lesen Sie dieses komplette Handbuch vor dem Verwenden des Instrumentes und verwenden Sie nur gem den Instruktionen Das Instrument muss immer mit der Erdleitung der Macht Schnur richtig niedergelegt zur Erde am Hauptausgang verwendet werden Nur unbesch digte elektrische Leitung und Ausr s tung spezifisch f r die Stromspannungen verwenden die Sie verwenden werden Die ganze mit der Hoch spannung verbundene Ausriistung sollte in Uberein stimmung mit EN61010 1 2001 sein Das Instrument ebenso trocken halten und reinigen wie m glich Wischen Sie regelmaBig mit einem weichen befeuchten Sie Stoff Lassen Sie d
3. Inc pourrait tre utilis pour fonctionner maintenir et entretenir ce produit Avertissant Parce que cet instrument peut d velopper la tension et le courant suffisants pour produire un choc mortel le soin doit tre exerc dans son op ration Cet instrument est conform ment con u a V EN61010 o piii 1 2001 norme de s curit lectrique N anmoins il devrait tre seulement utilis par les op rateurs convenablement entrain s Lire ce manuel entier avant utiliser l instrument et l usage seulement selon les instructions L instrument toujours doit tre utilis avec l avance de terre du cordon d alimentation correctement a fond a la terre a la sortie principale Utiliser le fil et l quipement lectriques seule ment intacts sp cifiques pour les tensions que vous utiliserez Tout quipement connect a haute tension devrait tre conform ment a EN61010 1 2001 Garder l instrument aussi sec et propre comme possi ble Essuyer r guli rement avec un doux touffer du tissu Laisser instrument seche compl tement avant l usage Ne pas fonctionner l instrument dans l extr me humidit au dessus de 80 Eviter la condensa tion en laissant l equilibrate d unit la temp rature ambiante en prenant l instrument d un plus froid un environnement plus chaud Permettre le refroidissement suffisant garantir que les conduits de l instrument ne sont pas couverts Wichtige Informationen
4. O rana Hoefer HE PLUS Horizontal Gel Electrophoresis System um HE PLUS Rev B0 07 12 Afro e fe r Contents Important Informatica ji Waste Electrical and Electronic Equipment WEE vii lef lee ix Operating Instructlons eee 1 Troubleshooting Guide 11 SOON and ne akan 12 RETOrENCES maama samma ries 13 Ordering Informati Oies nia 14 Care and Maintenance sussies isidir 15 Important Information English If this equipment is used in a manner not specified by Hoefer Inc the protection provided by the equipment may be impaired This instrument is designed for indoor laboratory use only Only accessories and parts approved or supplied by Hoefer Inc may be used for operating maintaining and servicing this product Warning Because this instrument can develop suffi cient voltage and current to produce a lethal shock care must be exercised in its operation This instrument is designed in accordance with the EN61010 1 2001 electrical safety standard Never theless it should be used only by properly trained operators Read this entire manual before using the instrument and use only according to the instructions The instrument must always be used with the earth lead of the power cord correctly grounded to earth at the mains outlet Use only undamaged electrical wire and equipment specific for the voltages you will use All equipment connected to high voltage should be in accordance wi
5. Plug the male ends of the black and red electrodes into the jacks on the side of the power supply After the samples have been loaded into the gel place the lid over the unit so that the lid covers align with the tank Set the lid straight down so that the lid rests squarely on the tank o Plug the power supply into a wall outlet F Setting the Power Supply Ensure an approved power cord that satisfies your regional voltage standard is used Input voltage is automatically detected by the system o Use the mode key gt to move between voltage amperage and time parameters To increase or decrease voltage use up A and down v arrow keys To increase or decrease amperage use up and down arrow keys o Set the timer Increase or decrease the value with the up and down arrow keys Between 99 hours and 59 minutes can be set as the run time Set 0 for Continuous On the left side of the Output key the flashing LED indicates that the timer operation has been paused When setting up the timer in this state set up after having pushed the output button for a long time so as to reset the timer Select the required output voltage up to 150 volt or 400 mA Press the start stop key to start the run CAUTION Do not jar or bump the gel box once the lid is place The electrical connection is made by gravity once the lid is in position While this design hel
6. rigtigt jordede til jord pa hovedledning sudlobet Bruger bare uskadt elektrisk trad og udstyr som v re specifik for spaendingerne du vil bruge Alt udstyr forbundet til haj spaending skulle vaere i overensstem melse med EN61010 1 2001 Beholder instrumentet s tor og ren som mulig Tor regul rt med et bl dt fugtigt stof Lad instrument O pii torken komplet for brug Driver ikke instrumentet i yderst fugtighed ovenfor 80 Undg kondensation ved lade enheden eguili brate til omgivende temperatur ved tageen instru mentets fra et koldere til et varmere miljo Attillade tilstraekkelig afkoling forsikrer at lufthullerne af instrumentet er ikke daekket Belangrijke Informatie Dutch Indien deze uitrusting in een manier wordt gebruikt die niet door Hoefer is gespecificeerd Nv de bescherming die door de uitrusting is verzorgd kan worden geschaad Dit instrument is voor binnenlaboratoriumgebruik enkel ontworpen Enkel onderdelen en delen keurden goed of leverden door Hoefer Nv kan voor het bedienen worden gebruikt handhavend en onderhouden van dit product Waarschuwend Omdat dit instrument voldoende spanning en stroom kan ontwikkelen om een dodeli jke schok te produceren moet zorg in zijn operatie worden geoefend Dit instrument is in overeenstemming met de EN61010 1 2001 elektrische veiligheidsstandaard ontworpen Niettemin zou het enkel door goed getrainde bedieningslieden moeten
7. set of 2 HE PLUS Mini Gel trays HE PLUS13 6 x 6 cm set of 4 HE PLUS Large Medium comb HE PLUS1428 14 28 Wells set of 2 HE PLUS Mini comb HE PLUSO508 5 8 Wells set of 2 HE PLUS Mini Casting kit HE PLUS16 Includes 4 gel trays 2 combs 5 8 Wells and casting stand HE PLUS Casting Stand for all HE PLUS17 3 gel sizes HE PLUS Standard Casting kit HE PLUS18 Includes 1 large gel tray 2 medium gel trays 4 combs 14 28 Wells and casting stand Important The units should never come into contact with the following cleaning agents these will cause irreversible and cumulative damage Acetone Phenol Chloroform Carbon tetrachloride Methanol Ethanol Isopropyl alcohol Alkalis Care and Maintenance Cleaning Units are best cleaned using warm water and a mild detergent Water at temperatures above 60 C can cause damage to the unit and components The units should not be left in detergents for more than 30 minutes The tank should be thoroughly rinsed with warm water and distilled water to prevent build up of salts but care should be taken not to damage the enclosed electrode Vigorous cleaning is not necessary or advised Air drying is recommended before use RNase Decontamination This can be performed using the following protocol e Clean the units with a mild detergent as described above e Wash with 3 hydrogen peroxide H202 for 10 minutes e Rinse with 0 1 DEPC diethyl pyrocarbonate treated distill
8. stand If you are using the 6 x 12 5 cm gels place the spacer in the center of the casting stand then insert the two 6 x 12 5 cm landscape gel trays Gel Divider When the gel solution has cooled to approximately 55 C slowly pour it into the gel tray If hotter gel solutions are routinely poured the tray may warp over time o If bubbles form on the surface of the gel upon pouring use the comb to either pop them or lightly brush them to the sides of the gel If large bubbles are allowed to harden within the gel they may cause artifacts to occur during electrophoresis mtn Insert one or more combs by placing them into the slots in the casting stand For best results place the comb in the slot nearest the end of the casting fixture If two combs are desired place the second in the center comb slot Comb Well Sample Description Width Vol 1 mm Code No 1 mm 14 tooth 5 mm 5 ul HE PLUS1428 dual format 1 mm 28 tooth 2 5 mm 2 5 pl HE PLUS1428 dual format 1 mm 5 tooth 8 mm 8 ul HE PLUSO508 dual format 1 mm 8 tooth 4 mm 4 ul HE PLUSO508 dual format Combs Allow the gel to harden undisturbed for at least 30 minutes Note Use of the same batch of electrophoresis buffer for both the gel and the running buffer is very important Slight variations in buffer composition between gel and running buffer may result in ionic or pH gradients that can significantly impact the mobility of the sampl
9. worden gebruikt Lees dit volledige handboek voor het gebruik het instrument en gebruik enkel volgens de instructies Het instrument moet altijd met de aardeleiding van het stroomsnoer correct grondde naar aarde aan het hoofdafzetgebied worden gebruikt Gebruik enkel onbeschadigde elektrische draad en uitrustings specifiek voor de spanningen u zult gebruiken Alle uitrustingen sloten aan aan hoogspan ning zou in overeenstemming met EN61010 1 2001 moeten zijn Houd het instrument zo droge en schone zoals mogelijk Bij Wis regelmatig met een zacht temper doek Verhuur het instrument droogt volledig voor het gebruik Bedien niet het instrument in extreme vochtigheid bovenstaande 80 Vermijd condensatie door het verhuren van de eenheid in evenwicht brengt naar omgevingstemperatuur wanneer nemen het instru ment van een kouder naar een lievere omgeving Om toe te staan voldoende afkoelen verzeker dat de luchtopeningen van het instrument niet bedekt zijn T rke Tietoa Finnish Jos t t varusteita k ytet n tavassa ei m ritetty Hoeferille Inc suojelu ehk isty varusteille saattaa olla avuton T m v line suunnitellaan sis laboratoriok yt lle vain Vain lis varusteet ja osat hyv ksyiv t tai toimitti Hoeferin oheen Inc voi k ytt k ytt miselle valvoalle ja servicing t m tuote Varoittaminen Koska t m v line voi kehitt riitt v j nnitteen ja virran t
10. 0 mM Tris base 20 mM Glacial Acetic Acid NaOAc 2 0 mM EDTA pH 8 3 10X Stock Solution 48 4 g Tris Base 16 4 g or 11 42 ml NaOAc 7 4 g EDTA or 20 ml 0 5 M EDTA pH 8 0 H20 to 1 liter Tris Borate EDTA Buffer TBE 1X Working Concentration 89 mM Tris Base 89 mM Boric Acid 2 0 mM EDTA pH 8 0 10X Stock Solution 108 g Tris Base 55 g Boric Acid 6 72 g EDTA or 40 ml 0 5 M EDTA pH 8 0 H20 to 1 liter Sample Loading Buffer DNA 6X Stock Solution 30 Glycerol in H20 0 25 Xylene cyanol 0 25 Bromophenol blue pH 8 References 1 Lehrach H et al 1977 Biochemistry 16 4743 Sambrook J Fritsch E F and Maniatis T 1989 Molecular Cloning A Laboratory Manual vol 1 Cold Spring Harbor Press New York Selden R E 1988 Analysis of RNA by Northern Hybridization in Current Proto cols in Molecular Biology EM Ausubel et al editors volume 1 p 4 9 1 Green Publish ing Associates and Wiley Interscience e pl3 e pl4 Ordering Information Product Code No HE PLUS Electrophoresis HE PLUS 115V complete system 115 V Includes gel tank safety lid power supply and standard casting kit HE PLUS Electrophoresis HE PLUS 230V complete system 230 V Includes gel tank safety lid power supply and standard casting kit Accessory Items HE PLUS Large Gel trays HE PLUS11 12 5 x 13 cm set of 2 HE PLUS Medium Gel trays HE PLUS12 12 5 x 6 cm
11. Permitir refrigeraci n suficiente asegure que las aberturas del Viktig Information Swedish om denna utrustning anv nds i ett s tt som inte har specificeras av Hoefer Inc skyddet tillhandah ll vid utrustningen kan skadas Detta instrument formges f r inomhuslaboratorium anv ndning bara Bara medhj lpare och delar godk nde eller levererade vid Hoefer Inc kan anv ndas f r fungera underh lla och servicing denna produkt varna D rf r att detta instrument kan utveckla tillr ck lig sp nning och str m att producera en d dlig st t m ste vas omsorg i dess funktion Detta instrument formges i verensst mmelse med EN61010 1 2001 elektriska s kerheten standarden Icke desto mindre bor det anv ndas bara av riktigt utbildade operat rer L s denna hela handbok f re anv nda instrumentet och anv nd bara enligt under visningarna Instrumentet m ste alltid anv ndas med jorden blyet av kraften repet riktigt grounded till jorden p det huvudutloppet Anv nder bara undamaged elektrisk tr d och utrust ning specifik f r sp nningarna du ska anv nda All utrustning kopplats som till h g spanning skulle vara i verensst mmelse med EN61010 1 2001 Haller instrumentet d torkar och reng r som m jlig Torka regelbundet med en mjuk fuktig trasa Lat instrumentet torka fullst ndigt f re anv ndningen Fungerar inte instrumentet i extrem fuktighet ver 80 Undvik kondenserin
12. as Instru ment trocken v llig vor dem Gebrauch Das Instrument in der u ersten Feuchtigkeit ber 80 nicht bedienen Vermeiden Sie Kondensation die Einheit equilibrate zur Umgebungstemperatur laBend wenn Sie das Instrument von einem k lteren bis eine w rmere Umgebung nehmen Um das gen gend Abk hlen zu erlauben stellen Sie sicher dass die Offnungen des Instrumentes nicht bedeckt werden Informazioni Importanti Italiano Se quest apparecchiatura usata in un modo specificato da Hoefer Inc la protezione fornito dall apparecchiatura potrebbe essere indebolita Questo strumento disegnato per l uso di laboratorio interno solo Solo gli accessori e le parti hanno approvato o hanno fornito da Hoefer Inc potrebbe essere usato per oper are per mantenere e per revisionare questo prodotto Avvertendo Perch questo strumento pu sviluppare il voltaggio sufficiente e la corrente di produrre una scossa letale la cura deve essere esercitata nella sua operazione Questo strumento disegnato conforme mente all EN61010 1 2001 la norma di sicurezza elettrica Tuttavia dovrebbe essere usato degli opera tori solo correttamente addestrati Leggere questo manuale intero prima di usare lo strumento e l uso solo secondo le istruzioni Lo strumento deve essere sempre usato col piombo di terra della spina di alimentazione correttamente hanno messo a terra alla terra alla presa di corrente principale
13. aven pro nap t budete pou vat V echna za zen spojen s vysok m nap t m by m la b t v souladu s EN61010 1 2001 Si ponech n stroje jako such a ist jako mo n Ot ete pravideln s a m kk vlhk m had kem Nech je n stroj nenastaven pln p ed pou it m Nejsou provozov na na n stroj v extr mn vlhkost nad 80 P ede lo kondenzaci o pron jmu jednotky na okoln teplotu nech p i p ij m n n stroj z chladn j do tepl ho prost ed Pro umo n n dostate n chlazen zajistit aby otvory n stroje jsou nevztahuje Vigtig Information Danish Hvis dette udstyr bruges i en made ikke specificeret ved Hoefer Inc den beskyttelse som er blevet forsynet af udstyret kan m ske svaekkes Dette instrument er designet for indendors laboratori umbrug bare Bare tilbeh r og del godkendede eller forsynede ved Hoefer Inc kan m ske bruges for drive funktionsfejl og betjening dette produkt Advare Fordi dette instrument kan udvikle tilstraek kelig spaending og strom at fremstille et dodbringende chok skal pleje bruges i dets drift Dette instrument er designet i overensstemmelse med EN61010 1 2001 elektrisk sikkerhedstandard Alligevel skulle det bruges bare af passende traenede operatorer Laes denne hel h ndbog for bruging instrumentet og brug bare i henhold til instruktionerne Instrumentet skal altid bruges med jordblyet af netledningen
14. cipe Allow the samples to migrate until the fragments have separated normally until the bromophenol blue dye front has migrated 34 of the way down the gel Note If the gel contains Ethidium Bromide the progress of electrophoresis may be monitored during the run by turning off the power supply removing the lid and shining a medium wave UV light onto the gel The resolved bands will appear as orange bands against a dark purple background o plo I Detection and Documentation of Separated Fragments 0 At the completion of the run turn off the power supply and disconnect the power cord Remove the lid and remove the gel tray Alternatively the entire tank can be placed on a Transilluminator o Ethidium Bromide stained samples are visualized by exposing them to medium wavelength 312 nm UV light Because the gel casting tray is UV transmittant the gel does not need to be removed from the tray before viewing Place the gel casting tray containing the gel on the filter surface of a UV Transilluminator for convenient viewing Troubleshooting Guide Problem Cause Solution The LCD screen remains blank and the fan does not run when the power is turned on Operation stops with alarm The screen displays NO LOAD AC power cord is not connected Check AC power cord connections at both ends Use the correct cords The fuse has blown Electrophoresis tank is not connected to the
15. dnie z tym EN61010 1 2001 Bezpiecze stwo elektryczne standard Niemniej jednak nale y stosowa jedynie przez odpowiednio przeszkoleni operator w Znajd pa stwo to ca y podr cznika przed zastosowaniem instrumentu i stosowa jedynie zgodnie z instrukcjami Instrument musi zawsze by wykorzystane z ziemi doprowadzi do zasilania detonuj cego w a ciwie uzasadnione na ziemi w sieci wodoci gowej rynku zbytu Wykorzystanie tylko nieuszkodzona elektrycznych drut w i urz dzenia specjalne do napi zap ac wykorzystania Wszystkie urz dzenia pod czone do wysokiego napi cia powinny by zgodne z EN61010 1 2001 Kontrolowa instrumentu jako suche i czyste jak to mo liwe Wytrze regularnie przy pomocy mi kkiego wilgotnej szmatki Niech si instrumentem ca kowicie wysuszy przed u yciem Nie prowadz do instrumentu w skrajnych wilgotno ci powy ej 80 Zapobiec kondensacji najmu przez jednostk r wnowa y do temperatury pokojowej przy podejmowaniu instrumentu z ch odniejsze w cieplejszych rodowiska Aby umo liwi wystarczaj ce ch odzenia zapewniaj e rozci cia of the instrument nie obj te ubezpieczeniem Informac es Importantes Portuguese Se este equipamento usado numa maneira n o espe cificada por Hoefer Inc que a protecc o fornecida pelo equipamento pode ser comprometida Este instrumento projectado para uso de interior de laborat rio s S aces
16. ed water e Caution DEPC is a suspected carcinogen Always wear gloves and safety glasses RNaseZAP Ambion can also be used Please consult the instructions for use with acrylic gel tanks e p15 Hoefer Inc 84 October Hill Road Holliston MA 01746 Toll Free 1 800 227 4750 Phone 1 508 893 8999 Fax 1 508 893 0176 E mail support hoeferinc com Web www hoeferinc com Hoefer is a registered trademark of Hoefer Inc RNaseZAP is a registered trademark of Ambion 2012 Hoefer Inc All rights reserved Printed in the USA Hoefer
17. es C Removing the Comb When the gel is solidified and fully opaque carefully remove the comb with a gentle wiggling upward motion If the comb is difficult to remove or if a low percentage gel is being used overlay the comb area with a small volume of 1X electrophoresis buffer to preserve the integrity of the wells Check the wells to ensure their bases are intact D Loading the Samples onto the Gel Remove the casting tray containing the hardened agarose gel from the casting fixture by lifting the ends Place the tray and gel into the main unit assembly such that the sample wells are on the same end as the negative black electrode e Fill the unit with the remaining 1X electrophoresis buffer containing Ethidium Bromide made previously covering the gel to a depth of 1 5 mm Approximately 350 ml of buffer will be required Load the samples into the wells with a micropipette or similar device taking care not to puncture the bottom of the wells or load the sample onto the top of the gel E Electrical Connections to the Safety Lid The Hoefer HE PLUS Gel System can only be operated with the safety lid in place Elec trical current is supplied through the tank electrodes from the power supply A micro switch connector in the power supply ensures a complete current path yet allows the lid to be removed from the unit without disturbing the loaded samples o Make sure the power supply is turned off a
18. g vid lata enheten equili brate till omgivande temperatur n r ta instrumentet fran en kallare till en varmare milj Att till ta tillr cklig kyla ser till att halen av instru mentet inte tacks o pvi English Ea French R German R Ea Italian R E Spanish La Swedish R Waste Electrical and Electronic Eguipment WEEE This symbol indicates that the waste of electrical and electronic eguipment 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 eguipment Ce symbole indique que les d chets relatifs a l quipement lectrique et lectronique ne doivent pas tre jet s comme les ordures m nag res non tri es et doivent tre collect s s par ment Contactez un repr sentant agr du fabricant pour obtenir des informations sur la mise au rebut de votre quipement Dieses Symbol kennzeichnet elektrische und elektronische Ger te die nicht mit dem gew hnlichen unsortierten Hausm ll entsorgt werden d rfen sondern separat behandelt werden m ssen Bitte nehmen Sie Kontakt mit einem autorisierten Beauftragten des Herstellers auf um Informationen hinsichtlich der Entsorgung Ihres Ger tes zu erhalten Questo simbolo indica che i rifiuti derivanti da apparecchiature elettriche ed elettroniche non devono essere smaltiti come rifiuti municipali
19. indifferenziati e devono invece essere raccolti separatamente Per informazioni relative alle modalit di smantellamento delle apparecchiature fuori uso contattare un rappresentante autorizzato del fabbricante Este s mbolo indica que el equipo el ctrico y electr nico no debe tirarse con los desechos dom sticos y debe tratarse por separado Contacte con el representante local del fabricante para obtener m s informaci n sobre la forma de desechar el equipo Denna symbol anger att elektriska och elektroniska utrustningar inte far avyttras som osorterat hushallsavfall och m ste samlas in separat Var god kontakta en auktoriserad tillverkarrepresentant f r information ang ende avyttring av utrustningen o pvii e pviii Packing List Hoefer HE PLUS Gel System Units include Tank with lid power supply power cord and standard casting set one 13 x 12 5 cm UV Transmittant Casting Tray two 6 x 12 5 cm UV Transmittant Casting Trays four 1 mm 14 28 reversible multichannel compatible combs and a casting stand The packing lists should be referred to as soon as the units are received to ensure that all components have been included The unit should be checked for damage when received save packaging for carrier Please contact your supplier if there are any problems or missing items Usage Guidance and Restrictions e Temperature range between 4 C and 40 C e Maximum relative humidity 80 for temperatures
20. l Concentrations and Resolving Ranges Concentration Agarose g Efficient Range of Agarose in per 100 ml of Separation Gel w V Buffer of Linear DNA Kb 0 3 0 3 5 60 0 6 0 6 1 20 0 7 0 7 0 8 10 0 9 0 9 0 5 7 1 2 1 2 0 4 6 L 1 5 0 2 3 2 0 2 0 0 1 2 Table taken from Sambrook J Fritsch E F amp Maniatis T 1989 Molecular Cloning A Laboratory Manual 1 6 8 613 Ps E Note The addition of Ethidium Bromide to both the gel and the running buffer will result in maximum detection levels by providing high levels of sample fluorescence with an evenly low level of background Make note of the total solution volume so that degree of evaporation can be determined and corrected for o Heat the agarose slurry in a microwave oven for 90 seconds Swirl the flask to make sure any grains sticking to the walls enter into the solution Undissolved agarose appears as small lenses floating in the solution Heat for an additional 30 60 seconds Re examine the solution and repeat the heating process until the agarose completely dissolves Add deionized water to replace any volume lost through evaporation during the heating process Add your detection reagent i e Ethidium Bromide to the manufacturers recommended concentration Mix by gently swirling the flask B Casting the Gel o Place the gel casting stand on a lab bench e Insert the gel casting tray into the casting
21. mentet og bruken bare gi til instruksjonene Instrumentet m alltid bli brukt med jorden blyet av kraftkabelen som riktig ha blitt jordet til jord p hovedledningen utlop Bruker bare uskadd elektrisk ledningsfremforing og utstyr som er spesifikk for spenningene du vil bruke All utstyr koplet til hoyspenning burde veere i samsvar med EN61010 1 2001 Beholder instrumentet som t rker og rengjor som mulig Visk regulaert med et mykt fuktig stoff La instru mentet torker komplett for bruk Driver instrumentet i ekstrem fuktighet ikke ovenfor 80 Unnga kondensasjon ved a la enheten equili brate til omgivelsestemperatur ved taen instrumentets fra et kaldere til et varmere miljo Til tillate tilstrekkelig kj lig sikrer at ventilasjon sapningene av instrumentet er ikke dekket Wazne Informacje Polish Je eli ten sprz t jest wykorzystywany w spos b nie okre lone przez Hoefer Inc do ochrony przewidzianej przez urz dzenie mo e zosta obni ony Instrument ten jest przeznaczony do u ytku w labora toriach kryty tylko Tylko akcesori w i cz ci zatwierdzone lub dostarczone przez Hoefer Inc mog by wykorzystane do eksp loatacji utrzymania i obs ugi tego produktu Uwaga Poniewa ten akt prawny mo e by rozwini cie odpowiednich napi cie i bie cych do wyprodu kowania miertelnego szoku opiek musi by wykony wane w dzia aniu Ten instrument zosta zaprojektowany zgo
22. power supply or there is a broken circuit in the electrophoresis cell Replace the fuse Check the connections to the power supply and on your electrophoresis cell to make sure the connection is intact check condition of wires in electrophoresis unit Close the circuit by reconnecting the cables Press RUN PAUSE to restart the run Buffer concentration incorrect Replace buffer Operation stops with alarm Display shows OVER VOLTAGE Circuit is interrupted Verify that the running buffer is correct Verify the all connections are attached correctly Turn the Power Switch off and on again restart application If you cannot restart the instrument turn off the power disconnect the power cord from the outlet and contact Technical Service Operation stops with alarm Display shows LEAKAGE Ground leak detected during run Check the electrophoresis system for improper grounding Restart the power supply by turning the Power switch off and on Turn power off then check the gel tank for buffer leakage LID alarm Cover of gel tank not in correct place Turn off power supply Place the lid so the magnet is pressed against the power supply and restart If you cannot restart the instrument turn off the power disconnect the power cord from the outlet and contact Technical Service e pll o pl2 Solutions Tris Acetate EDTA Buffer TAE 1X Working Concentration 4
23. ps to minimize sample disturbance during lid placement it also may result in a disruption of power to the unit if the lid or unit are disturbed during the run CAUTION DO NOT EXCEED THE MAXIMUM OPERATING VOLTAGE OF 150 VOLTS G To Pause a Run and or Change Parameters o To pause the run select Pause During the pause mode the voltage amperage or time can be changed by highlighting the function and using the arrow keys then pressing enter Once the changes have been made the start button can be pressed to resume the run e To stop the run press the pause button for 30 seconds Stop will appear H Sample Electrophoresis 0 The maximum suggested applied voltage for the electrophoresis of DNA in agarose gels using the Hoefer HE PLUS Gel System is 150 volts e Ina 1 TBE gel this translates into a run time of approximately 1 hour Lower voltages may be used of course and as a general rule a 70 V run will take twice as long as a 145 V run Higher voltages may be used to decrease run time however if the unit is being operated at higher voltages than 150 V the heat generated during electrophoresis may decrease sample resolution Such artifacts may be avoided by running the unit in a cold room or adding 1X electrophoresis buffer ice cubes to keep the unit properly cooled Follow the sample migration into the gel using the loading dye as an indicator See page 12 for the Sample Loading Buffer re
24. s rios e partes aprovaram ou forneceu por Hoefer Inc pode ser usada para operar Manter e servicing este produto Advertindo Porque este instrumento pode desen volver voltagem suficiente e corrente produzir um choque letal cuidado deve ser exercitado em o pv sua Operac o Este instrumento projectado de acordo com o EN61010 1 2001 condic o de seguranca el ctrica Nao obstante deve ser usado s por operadores adequada mente treinados Leia este manual inteiro antes de usar o instrumento e use s de acordo com as instrucc es O instrumento sempre deve ser usado com o chumbo de terra do cord o de poder corretamente baseou a terra nos canos sa da principais Usa fio el ctrico s intacto e equipamento espec fico para as voltagens que voc usar Todo equipamento conectado a voltagem alta deve ser de acordo com EN61010 1 2001 Mantem o instrumento tao seco e limpo como possivel Limpe regularmente com um pano h mido macio Deixe o instrumento secar completamente antes de uso Nao opera o instrumento em humidade extrema acima de 80 Evite condensac o deixando o equili brate de unidade a temperatura ambiental quando tomar o instrumento de um mais frio a um ambiente mais quente Permitir esfriar suficiente assegura que as aberturas do instrumento n o s o cobertas Informaci n Importante Spanish Si este equipo es utilizado en una manera no especifi cado por Hoefer S a la pro
25. s for Selecting Electrophoresis Buffers and Gel Concentrations The two most commonly used buffers for hori zontal electrophoresis of double stranded DNA in agarose gels are Tris Acetate EDTA TAE and Tris Borate EDTA TBE While the resolv ing powers of these buffers are very similar the relative buffer capacities are very different conferring different run attributes which are summarized below TAE Tris acetate has traditionally been the more commonly used buffer However ts relatively low buffer capacity will become exhausted during extended electrophoresis making buffer recirculation necessary in runs exceeding 140 mA hours Potential advantages of using TAE buffer over TBE buffer include superior resolution of supercoiled DNA and approximately 10 faster migration of double stranded linear DNA fragments TBE Tris borate s significantly greater buffering capacity and its relatively low current draw eliminates the need for recirculation in all but the most extended runs gt 300 mA hours TBE buffer systems are not recommended when fragments are to be recovered from the gel after electrophoresis Ps O Make 500 ml of either 1X TAE or 1X TBE electrophoresis buffer 9 Weigh an appropriate quantity of agarose see Table 1 and place it into a 250 ml flask Add a sufficient quantity of either 1X TAE or 1X TBE buffer prepared in step 1 to achieve a final volume of 100 ml agarose solution Table 1 Ge
26. tecci n proporcionado por el equipo puede ser da ada Este instrumento es dise ado para el uso interior del laboratorio s lo S lo accesorios y partes aprobaron o suministraron por Hoefer S a puede ser utilizado para Operar para mantener y para atender a este producto Advirtiendo Porque este instrumento puede desar rollar voltaje y corriente suficientes para producir un golpe mortal el cuidado debe ser ejercitado en su operaci n Este instrumento es dise ado de acuerdo con el EN61010 1 2001 est ndar el ctrico de seguridad No obstante debe ser utilizado s lo por operarios adecuadamente capacitados Lea este manual entero antes de utilizar el instrumento y el uso s lo seg n las instrucciones El instrumento siempre debe ser utilizado con el plomo de la tierra del cable de alimentaci n moli correcta mente a la tierra en la salida de red Utiliza alambre y equipo el ctricos s lo ilesos espec ficos para los voltajes que usted utilizar Todo equipo conectado al voltaje alto debe ser de acuerdo con EN61010 1 2001 Mantiene el instrumento tan seco y limpio como posible Enjugue regularmente con un suave el trapo h medo Permita que el instrumento seque completa mente antes de uso No opera el instrumento en la humedad extrema encima de 80 Evite condensaci n permitiendo la unidad equilibra a la temperatura ambiente al tomar el instrumento de un m s fr o a un ambiente m s tibio
27. th EN61010 1 2001 Keep the instrument as dry and clean as possible Wipe regularly with a soft damp cloth Let the instrument dry completely before use Do not operate the instrument in extreme humidity above 80 Avoid condensation by letting the unit equilibrate to ambient temperature when taking the instrument from a colder to a warmer environment To permit sufficient cooling ensure that the vents of the instrument are not covered Dulezit Informace Czech Pokud by toto za zen je pou ito zp sobem kter neni podle Hoefer ochrana poskytovana na zaklad Inc za zen m e byt naru ena Tento n stroj je ur en pro vnit n pou it v laborato i pouze Pouze p slu enstv a sti schv len nebo poskyt nut ch Hoefer Inc mohou b t pou ity pro provoz dr bu a dr b tohoto v robku Pozor Proto e tento n stroj m e vyvinout dostate n nap t a proud kter m vyr b t a smrtiacej ok p e mus b t vykon v na v jeho provoz Tento n stroj je ur en v souladu s EN61010 1 2001 elektrick bezpe nostn normy P esto e by m ly b t pou ity pouze dn vy kolen oper tor st cel toto ru n p ed pou it m n stroje a pou it pouze v souladu s pokyny P stroj mus b t v dy pou vaj se na v konu zemi v st ra spr vn zemn ny k zemi na s ti v stce Vyu it pouze nepo kozen elektrick dr ty a vyb
28. uottaa kuolettavan j rkytyksen huolta t ytyy harjoittaa toiminnossaan T m v line suunnitellaan EN61010 1 2001 s hk turvallisuusstandardin mukaisesti Silti pit isi k yt t vain ohi oikeasti koulutetut k ytt j t Lue t m kokonainen manuaalinen ennen v linett ja k ytt vain ohjeiden mukaan V linett t ytyy k ytt aina valtanuoran maalyijyst perusti oikein maadoittaa s hk verkkoaukossa K ytt vain undamaged s hk metallilankaa ja varus teita t sm llinen j nnitteille k ytt Kaikki varusteet yhdistetty korkeaan j nnitteeseen pit isi olla EN61010 1 2001IN mukaisesti Pit v lineen yht kuiva ja puhdas kuin mahdollinen Pyyhi s nn llisesti pehme ll kostealla kankaalla Anna v line kuivua t ysin ennen k ytt Ei k yt v linett extreme ilmankosteudessa 80 n yl puolella V lt tiivistymist antamalla yksik n equili brate ymp r iv n l mp tilaan kun ottaminen v line kylmempi l mpim mp n ymp rist n Sallia riitt v n j hdytt minen varmistaa ett v lineen ilmarei t peitet n Information Importante French Si cet quipement est utilis dans une mani re pas sp cifi par Hoefer Inc la protection fourni par V guipement pourrait tre diminu e Cet instrument est con u pour l usage de laboratoire int rieur seulement Seulement les accessoires et les parties ont approuv ou ont fourni par Hoefer
29. up to 31 C decreasing linearly to 50 relative humidity at 40 C e Not for outdoor use This apparatus is rated POLLUTION DEGREE 2 in accordance with IEC 664 POLLUTION DEGREE 2 states that Normally only non conductive pollution occurs Occasionally however a temporary conductiv ity caused by condensation must be expected Specifications Unit dimensions W x L x H Gel dimensions W x L Maximum sample capacity Buffer capacity Distance between electrodes Electrophoresis Tank Overall dimension W x L x H Material characteristic Solution volume Safety Lid Overall dimension W x L x H Material characteristic Power Supply Overall dimension W x L x H Weigh Input voltage Output voltage Output amperage Maximum wattage Timer Safety switch Memory function 24 5 x 17 0 x 6 2 cm 12 5 x 13 0 cm 112 samples 4 combs 28 samples each 350 ml 13 5 cm 18 3 x 16 4 x 5 6 cm UV transmitting 50 at 254 nm 80 at 312 nm 350 ml includes buffer and gels 19 7 x 16 9 x 3 8 cm UV non transmitting acrylic 7 5 x 17 0 x 6 2 cm 410 g AC 100 240 V 50 60 Hz 10 to 150 volts in 10 V steps Constant peak voltage of 150 V 10 to 400 mA in 5 mA steps 45 W 99 hours 59 min and continuous model A microsensor in the power supply prevents output without the safety lid in place Automatic memory the last used Volt and Time o pix Operating Instructions A Guideline
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