Hoefer HE99X User Manual
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Hoefer HE99X User Manual

Submarine electrophoresis unit
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Hoefer HE99X
Submarine electrophoresis unit
um
HE99X-IM/Rev. L0/07-12

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  • Page 1 Hoefer HE99X Submarine electrophoresis unit HE99X-IM/Rev. L0/07-12...

  • Page 2: Table Of Contents

    Contents   Important Information   ........... ii Waste Electrical and   Electronic Equipment (WEEE) .......vii Submarine Electrophoresis Unit   Function and description  ........1 Specifications............2 Operating instructions  ..........3 Care and maintenance ...........8 Troubleshooting .............9 Buffers, volumes, and notes .........10 Loading buffer and sample volumes ......12 Ordering information  ..........16 •  pi...

  • Page 3: Important Information

    Important Information – English před odběrem energie vede bezpečnostní víko. • Rozeslat pouze voda nebo 50/50 voda/ • If this equipment is used in a manner not specified ethylenglykolu prostřednictvím výměník tepla je by Hoefer, Inc. the protection provided by the li to vybavena. Nemají připojení výměník tepla s equipment may be impaired. vodními setřepná nebo jakékoli chladicí kapaliny • This instrument is designed for indoor laboratory zdroje, kde tlak vody je neregulo. use only. • Nikdy zavést prostředek proti zamrznutí nebo • Only accessories and parts approved or supplied jakákoli organická rozpouštědla do jakékoli by Hoefer, Inc. may be used for operating, části z tohoto nástroje. Rozpustidlům způsobí maintaining, and servicing this product. nenapravitelné poškození jednotka! • Only use a power supply that is CE marked or • Nejsou provozována s pufru teplotách nad safety certified by a nationally recognized testing maximální stanovenou technickými specifikacemi. laboratory. Přehřátí způsobí nenapravitelné poškození • The safety lid must be in place before connecting jednotka! the power supply leads to a power supply. Vigtig Information – Danish • Turn all power supply controls off and disconnect...
  • Page 4 Belangrijke Informatie – Dutch joka on kansallisesti tunnustettnut testaaminen laboratoriota. • Indien deze uitrusting in een manier wordt • Turvallisuuskansi täytyy olla paikallaan gebruikt die niet door Hoefer, Inc. is gespecificeerd ennen yhdistäminen käyttöjännitelyijyjä de bescherming die door de uitrusting is verzorgd käyttöjännitteeseen. kan worden geschaad. • Kiertää kaikki käyttöjännitevalvonnat ja irrottaa • Dit instrument is voor binnenlaboratoriumgebruik valtalyijyt ennen poistaminen turvallisuuskantta. enkel ontworpen. • Kiertää vain vesi tai 50/50 vesi/ethyleneä glycol • Enkel onderdelen en delen keurden goed of siinä tapauksessa varustetun lämmönvaihtimen leverden door Hoefer, Inc. kan voor het bedienen läpi. Älä yhdistä lämmönvaihdinta worden gebruikt, handhavend en onderhouden vesinapautukseen eikä jäähdytysnestelähteeseen, van dit product. missä vesipaine on unregulated. • gebruik Enkel een netvoeding die CE is markeerde • Pakkasneste eikä orgaaninen liuotin välineen of veiligheid die door een is gecertificeerd die osassa ei esitele Koskaan. Orgaaniset liuottimet nationaal is herkend testene laboratorium. aiheuttavat korvaamattoman vahingon yksikköön! • Het veiligheidsdeksel moet in plaats voor het...
  • Page 5 Informazioni Importanti – Italian l’instrument. Les dissolvants organiques causeront des dommages irréparables à l’unité! • Se quest’apparecchiatura è usata in un modo • Ne pas fonctionner avec les températures de specificato da Hoefer, Inc. la protezione fornito tampon au-dessus du maximum a spécifié des dall’apparecchiatura potrebbe essere indebolita. spécifications techniques. La surchauffe causera • Questo strumento è disegnato per l’uso di des dommages irréparables à l’unité ! laboratorio interno solo. • Solo gli accessori e le parti hanno approvato o Wichtige Informationen – German hanno fornito da Hoefer, Inc. potrebbe essere • Wenn diese Ausrüstung gewissermaßen nicht usato per operare, per mantenere, e per revisionare angegeben durch Hoefer, Inc. verwendet wird, questo prodotto. kann der durch die Ausrüstung zur Verfügung • usa Solo un alimentatore che è CE ha marcato gestellte Schutz verschlechtert werden. o la sicurezza certificato da un nazionalmente • Dieses Instrument wird für den riconosciuto testando il laboratorio. Innenlaborgebrauch nur dafür entworfen. • Il coperchio di sicurezza deve essere nel luogo • Nur Zusätze und Teile genehmigten oder lieferten...
  • Page 6 Przegrzania spowoduje nieodwracalne szkody dla utstyrer. Ikke forbind oppvarmingen veksleren jednostki! til en vanntapp eller noe kjølemiddelkilde hvor vannet trykket er unregulated. Informações Importantes – • Introduserer Aldri antifreeze eller noe organisk Portuguese løsemiddel inn i noe del av instrumentet. Organiske løsemiddler vil forårsake irreparabel • Se este equipamento é usado numa maneira skade på enheten ! não especificada por Hoefer, Inc. que a • Driver med buffertemperaturer over maksimum protecção fornecida pelo equipamento pode ser ikke spesifiserte teknisk spesifikasjoner. Å comprometida. overoppheting vil forårsake irreparabel skade på • Este instrumento é projectado para uso de interior enheten ! de laboratório só. • Só acessórios e partes aprovaram ou forneceu por Wazne Informacje – Polish Hoefer, Inc. pode ser usada para operar, manter, e • Jeżeli ten sprzęt jest wykorzystywany w sposób servicing este produto. nie określone przez Hoefer, Inc. do ochrony • Só usa um estoque de poder que é CE marcou...
  • Page 7 Información Importante – Spanish • Säkerheten locket måste vara på platsen före koppla kraften tillgången blyen till en kraft tillgång. • Si este equipo es utilizado en una manera no • Vänder sig alla kraft tillgång kontroller av och especificado por Hoefer, Inc. la protección kopplar bort kraften blyen före flytta säkerheten proporcionado por el equipo puede ser dañada. locket. • Este instrumento es diseñado para el uso interior • Cirkulerar bara vatten eller 50/50 vatten/ethylene del laboratorio sólo. glycol genom värmen exchanger i så utrustad fall. • Sólo accesorios y partes aprobaron o suministraron Inte kopplar värmen exchanger till en vatten kran por Hoefer, Inc. puede ser utilizado para operar, eller något kylmedel källa där vattnet trycket är para mantener, y para atender a este producto. unregulated. • Sólo utiliza una alimentación que es CE marcó o • Inför aldrig kylvätska eller något organiska la seguridad certificada por un nacionalmente lösningsmedel in i någon del av instrumentet. reconocido probando el laboratorio. Organiskt lösningsmedel ska orsaka irreparable • La tapa de la seguridad debe estar en el lugar skada till enheten! antes de conectar la alimentación lleva a una • Använd inte med buffert temperaturer över...

  • Page 8: Waste Electrical And Electronic Equipment (Weee)

    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. Ce symbole indique que les déchets relatifs à l’équipement  French électrique et électronique ne doivent pas être jetés comme  les ordures ménagères non-triées et doivent être collectés  séparément. Contactez un représentant agréé du fabricant  pour obtenir des informations sur la mise au rebut de  votre équipement. German Dieses Symbol kennzeichnet elektrische und elektronische  Geräte, die nicht mit dem gewöhnlichen, unsortierten  Hausmüll entsorgt werden dürfen, sondern separat  behandelt werden müssen. Bitte nehmen Sie Kontakt mit  einem autorisierten Beauftragten des Herstellers auf, um  Informationen hinsichtlich der Entsorgung Ihres Gerätes  zu erhalten.  Italian Questo simbolo indica che i rifiuti derivanti da  apparecchiature elettriche ed elettroniche non devono essere  smaltiti come rifiuti municipali indifferenziati e devono invece  essere raccolti separatamente. Per informazioni relative alle  modalità di smantellamento delle apparecchiature fuori uso,  contattare un rappresentante autorizzato del fabbricante.       Spanish Este símbolo indica que el equipo eléctrico y electrónico no  debe tirarse con los desechos domésticos y debe tratarse por  separado. Contacte con el representante local del fabricante ...

  • Page 9: Submarine Electrophoresis Unit Function And Description

    Submarine Electrophoresis Unit Function and description ® The Hoefer HE99X unit electrophoretically separates nucleic acid fragments in a submarine gel. The gel is first cast in a gel caster, which is available in three lengths. Once the gel sets, the...

  • Page 10: Specifications

    If any part is missing, contact your local Hoefer, Inc. sales office. Inspect all components for damage that may have occurred while the unit was in transit. If any part appears damaged, contact the carrier immediately.

  • Page 11: Operating Instructions

    Operating instructions Before you start… Agarose gels are first cast in the gel casting kit, and samples are then loaded into the wells and 1.   W ash all components  with a dilute solution of  electrophoretically separated. The fluorescent laboratory detergent and rinse  dye ethidium bromide can be added to the gel thoroughly.
  • Page 12 Prepare the casting tray and pour the gel Install a foam pad at each end of the casting tray. Use a comb as a placement guide so that the pad  adheres  ~ ~ 1 mm from the bottom of the tray: Lay  the comb into the bottom of the tray, oriented so  that it fits completely across the tray along the side  that is 16 cm wide. Peel off the adhesive backing on  the foam pad, align the pad on the comb, adhesive  side toward the inside wall of the tray, and slide the  comb against the wall. Press the foam pad in place  and repeat with second pad on the wall opposite the  first pad. Color-coded dots (2)  Fig 2. Gel casting kit. on handles orient the  Running tray installation: Approach  running tray on the ...
  • Page 13 Seat the running tray between the foam pads in the  casting tray by placing one end of the tray against  the foam pad, slightly compressing it, then seating  the other end of the tray against the opposite foam  pad. (See arrows A and B in Fig 2.) The running tray  should lay flush against the bottom of the casting tray. Place the casting tray assembly on a leveling surface  and level, using the spirit level on the running tray as  a guide. Check that the comb assembly leaves  ~ ~ 1 mm  of space between the comb bottom and the running  tray. Remove the level and the comb assembly. Prepare the combs Align the two slots in the comb with the loosened  Fig 3. Assembled comb. thumb screws of the comb back. Tighten the screws  until the comb is just supported.  comb back Place the comb assembly into a set of slots on the  running tray seated in the casting tray. Adjust the  screws comb so that the bottom of the teeth are  ~ ~ 1.0 mm  from the running tray. Tighten the screws to secure  the comb.  To run twice as many samples on the 15 and 20 cm  trays, prepare two comb assemblies and place one  near the cathode end, indicated by the black dot, and  one at the center.  •  p5...
  • Page 14 Final casting steps Pour the agarose solution (cooled to 50 °C) onto the  running tray seated in the casting tray. Orient the  comb assembly so that it is at the end of the tray  opposite the direction of migration (typically at the  cathode [–] end, which is marked by a black dot on  the handle). Fit the comb assembly into the slots. Allow a minimum of 30 min for the gel to set, then  remove the comb carefully: partially lift and slightly  tilt the comb at one end and slowly withdraw it from  the gel. (Pulling the comb straight up creates a  vacuum in the wells that may lift the gel out of   the tray.) Lift the running tray out of the casting tray and  transfer it with the gel to the horizontal unit. Orient  the running platform so that the sample will “run to  red”. That is, place the sample wells at the cathode  (–) end, which is indicated by a black dot. A notch on  either side of the running tray centers the tray on the  running platform.  •  p6...
  • Page 15 Preparing for electrophoresis Optional: To monitor separation progress, either add  0.5 µg/ml (final conc.) of ethidium bromide to the  running buffer now or add 50 µg/ml (final conc.)  Caution! Wear UV safety goggles  ethidium bromide to the sample buffer. To visualize  and protect skin when using a  progress, turn off the power supply, remove the lid  UV lamp. assembly, and hold a portable UV lamp near the gel.   Note: Adding ethidium bromide to the running or  Note: Refer to the Buffers,  sample buffer slows migration slightly. Detection by  volumes, and notes section  this method is not as sensitive as by staining after the  for additional information and  electrophoresis run. See the DNA detection section,  guidelines on page 10. for more details (page 14).  See page 12 for a sample  loading buffer recipe and well  volumes for various comb sizes  Fill the chamber with buffer until the gel is  in gels of different thicknesses. submerged  ~ ~ 1 mm. Load the samples. Add the sample to 1/5 volume of  the sample loading buffer. Mix each sample and load  into a well with a micro-pipet, taking care to avoid  Important!  If running two sets  puncturing the well bottom or entrapping bubbles. ...

  • Page 16: Care And Maintenance

    After electrophoresis Important!  Always turn off the power supply and  disconnect the leads before removing the lid. If no ethidium bromide was added to the gel or  sample before the run, stain the gel now in a solution  of 0.5 to 1.0 µg/ml ethidium bromide in water  or buffer. Clean the unit as described in the next section. Care and maintenance Cleaning • Never autoclave or heat any component above 45 °C. • Never use abrasive cleansers. • Do not expose the unit to solutions or vapors of aromatic or halogenated hydrocarbons, ketones, esters, alcohols (over 30%), or concentrated acids (over 25%).

  • Page 17: Troubleshooting

    Troubleshooting problem solution Sample well deformed    A llow the gel to set for a minimum of 1 hour and make  sure it is at room temperature before removing the comb.     R emove the comb at a slight angle and very slowly to  prevent the gel from breaking.     T ake care to not damage the well with the pipet while  loading the sample; aim for the center of the well and do  not puncture the bottom with the pipet tip. Samples not running   If the comb is warped, replace. along a straight path     I f the running tray is warped, replace.  (Cool agarose to 50 °C to prevent the tray from warping.)     C irculate buffer if it becomes depleted by stopping   the run and pipetting the buffer from one chamber to   the other. ...

  • Page 18: Buffers, Volumes, And Notes

    Notes, buffers, and volumes Running buffers for DNA in agarose gels Recipes for the three most commonly used running buffers for DNA electrophoresis are listed below. The buffering capacity of both Important!  Do not adjust the  pH of these buffers once   TBE and TPE is usually sufficient so that buffer they are prepared according  ...
  • Page 19 2. 10X Tris-phosphate-EDTA (TPE) stock buffer (0.89 M Tris, 0.89 M phosphoric acid, 20 mM EDTA, pH ≈8.1, 1000 ml) Tris base (FW 121.1)  0.89 M  108.0 g Phosphoric acid (85%)  0.23 M  15.5 ml EDTA solution  0.02 M  40.0 ml  (0.5 M, pH 8.0, solution 4) Deionized H O     to 1000.0 ml Stir. Do not adjust pH. Dilute to 1X, to yield 89 mM  Tris base, 23 mM phosphoric acid, and 2 mM EDTA.  3. 10X Tris-acetate-EDTA (TAE) stock buffer (0.4 M Tris, 0.2 M acetic acid, 10 mM EDTA, pH ≈8.4, 1000 ml) Tris base (FW 121.1) ...

  • Page 20: Loading Buffer And Sample Volumes

    Loading buffer and sample volumes Loading buffer (5X, 25% Ficoll 400, 0.25% Bromphenol blue † , 10 ml) Deionized H O     to 7.0 ml Ficoll 400     2.5 g Bromophenol blue (FW 691.9)    25.0 mg Deionized H O     to 10.0 ml Add 1 volume loading buffer to 4 volumes of sample.  (Loading buffer increases solution density.) Note 1:    S ucrose or glycerol may be used instead of  Ficoll 400. Note 2: ...
  • Page 21 Agarose gel electrophoresis notes Agarose gel electrophoresis can be used to separate DNA fragments down to 0.1 kb or less. Polyacrylamide gels are typically used for fragments smaller than 1 kb. DNA mobility The suggested agarose concentration for separating fragments of various sizes is listed below.
  • Page 22 RNA samples usually require longer runs or buffers that are easily depleted, and so require circulation. The Hoefer SUB20C and SUB25C horizontal units are recommended for this appli- cation rather than the HE33. DNA detection Caution! Ethidium bromide is ...
  • Page 23 place the gel on the transilluminator, ensure that it lies flat by cutting off the ridges formed by the grooves in the running tray. (Do not damage the transilluminator surface; trim both ends of the gel with a spatula while it is still in the tray, lift away the ridges, and then slide the gel onto the transilluminator.) For viewing, 302 nm light is recommended for both acceptable sensitivity and...

  • Page 24: Ordering Information

    4  HE98X Leveling feet  4  HE99XRK-1 Companion products MacroVue ™  UV-20 Transilluminator 115 V~  1  UV20-115V MacroVue ™  UV-20 Transilluminator 230 V~  1  UV20-230V HE99X gel casting kits Each size includes 1 gel casting tray, 1 UVT running tray, and 4 foam sealing gaskets 15 × 10 cm  1  HE97X-10 15 × 15 cm  1  HE97X-15 15 × 20 cm  1  HE97X-20 HE99X gel casting trays 15 × 10 cm  1  HE95X-10 15 × 15 cm  1 ...
  • Page 25 Combs comb thickness well width no. of wells (mm) (mm) code number 1/2*  1.5  113/10  HE91A-P-1.5 1/2*  3.0  113/10  HE91A-P-3.0 10  1.5  9.7  HE91A-10-1.5 10  3.0  9.7  HE91A-10-3.0 15  1.0  7.1  HE91A-15-1.0 15  1.5  7.1  HE91A-15-1.5 15  3.0  7.1  HE91A-15-3.0 20 ...
  • Page 26 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.  © 2012 Hoefer, Inc. —   All rights reserved. Printed in the USA.

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