C.B.S. Scientific DH User Manual

INSTRUCTION MANUAL
Dedicated Height Nucleic Acid
Sequencing Systems
Single Unit Dual Unit
DH-400-20 DDH-400-20 DH-500-20 DDH-500-20 DH-600-20 DDH-600-20 DH-400-33 DDH-400-33 DH-500-33 DDH-500-33 DH-600-33 DDH-600-33
TABLE OF CONTENTS
Important User Information 3-4 Section 1 General Information
1.1 Introduction 5
1.2 Specifications 5
1.3 Safety 6
Section 2 Description of parts
2.1 Unpacking-Dedicated Height Sequencer 7
2.2 Unpacking-Dual Dedicated Height Sequencer 8
Section 3 Assembly
3.1 Unit assembly for Single Dedicated Sequencer 9-10
3.2 Unit assembly for Dual Dedicated Sequencer 10
3.3 Glass Plate Preparation 11
3.4 Methods for Sealing Glass Plate Sandwich 12-13
3.5 Pouring the Gel 14-15
Section 4 Electrophoresis
4.1 Pre-Electrophoresis 16-17
4.2 Loading Samples 17
4.3 Electrophoresis 18
4.4 Post Electrophoresis-Gel Processing 19-20
4.5 Recommended Buffers and Reagents 21
4.6 References and Suggested Reference Literature 22
Section 5 Tables: Power Settings, Troubleshooting, Comb Volumes
5.1 Recommended Power Settings 23
5.2 Troubleshooting Guide 24
5.3 Comb/Well Volumes 25-27
Section 6 Maintenance of Equipment
6.1 Care and Handling 28
6.2 Maintenance 28
Section 7 Ordering Information 29-37
C.B.S. Scientific 2 Dedicated Height Sequencer
S
ENGLI
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IMPORTANT USER INFORMATION
This Instruction Manual will explain how to use this product safely and effectively. Please read and carefully follow the instruction manual in its entirety.
The triangle/exclamation mark symbol alerts the user of the product to important operational, maintenance, and/or warranty requirements.
The triangle/lighting bolt symbol alerts the user of the product to potentially hazardous
WARRANTY AND LIABILITY
This product was produced utilizing the highest practical standards of materials, workmanship, and design. C.B.S. Scientific warrants that the product has been tested and will meet or exceed published specifications. This warranty is valid only if the product has been operated and maintained according to the instructions provided.
C.B.S. Scientific warrants this product to be free from defects in materials and workmanship under normal service for one year from date of shipment. If the product proves defective during this period, C.B.S. Scientific will repair or replace it at our option, free of charge, if returned to us postage prepaid. This warranty does not cover: damage in transit, damage caused by carelessness, misuse or neglect, normal wear through frequent use, damage caused by solvent corrosion, damage caused by improper handling or user alteration, nor unsatisfactory performance as a result of conditions beyond our control. C.B.S. Scientific shall in no event be liable for incidental nor consequential damages, including without limitation, lost profits, loss of income, loss of business opportunities, loss of use and other related damages, however caused, nor any damage arising from the incorrect use of the product.
electrical exposure.
Failure to adhere to the instructions could result in personal and/or laboratory hazards, as well as invalidate any warranty. Always turn off the DC power source prior to disconnecting power cords from the product. Disconnect power cords from the power source first, and then from the product. For maximum safety, always operate this system in an isolated, low traffic area, not accessible to unauthorized personnel. Never operate damaged or leaking equipment.
C.B.S. Scientific 3 Dedicated Height Sequencer
FRANÇAIS INFORMATION IMPORTANTE À L'USAGE DES UTILISATEURS
Le présent manuel d'utilisation explique la manière de se servir efficacement du produit en conditions
de sécurité. Il est recommandé de soigneusement lire la totalité du manuel, avec ses consignes et ses instructions.
Le triangle avec point d'exclamation est un symbole destiné à avertir l'utilisateur du produit de
l'importance de certaines exigences relatives au fonctionnement, à l'entretien et/ou à la garantie.
Le triangle avec flèche en zigzag est un symbole destiné à avertir l'utilisateur du produit de la possibilité d'exposition à des décharges avec danger de secousses électriques.
Tout manquement à l'observation des consignes et des instruc tions peut exposer les personnes et les biens à des dommages corporels et/ou m atériels et peut annuler toute garantie. Il faut toujours interrompre l'alimentation de courant continu avant de déconnecter les cordons d'alimentation du produit. Déconnecter d'abord les cordons d'alimentation branchés sur la source de tension (alimentation de secteur) puis ceux branchés sur le produit. Pour une sécurité m aximum, il faut toujours faire fonctionner ce système dans un lieu isolé, peu fréquenté, où le personnel non autorisé n'a pas accès. Ne jamais faire fonctionner un matériel endommagé ou affecté par des fuites.
GARANTIE ET RESPONSABILITÉ
Le produit a été fabriqué conformément aux norm es applicables les plus exigentes en matière de matériaux, de main d'oeuvre, de conception et d'ingéniérie. C.B.S. Scientific garantit que le produit a subi des essais et que ses performances r empliront les conditions des spéc ifications publiées ou leur seront même supérieures. La présente garantie n'est valide que si le produit a fonctionné et a été entretenu conformément aux consignes et instructions fournies.
C.B.S. Scientific garantit que le produit sera dépourvu de vices de matériaux et de main d'oeuvre, en conditions de service normales, pendant un an à com pter de la date d'expédition. Au cas où le pr oduit s'avérerait défectueux pendant cette période de garantie, C.B.S. Scientific réparera ou r emplacera le produit, à sa discrétion et gratuitement, si le produit lui est retourné port payé d'avance. La garantie ne couvre pas les dommages de transport; les dommages causés par l'im prudence, le manque de soins, l'abus ou la négligence; l'usure normale résultant d'une utilisation fréquente; les domm ages causés par la corrosion des solvants; et les dommages causés par la manipulation inadéquate ou des changements apportés par l'utilisateur. La garantie ne couvre pas non plus les perf ormances non satisfaisantes résultant de conditions hors du contrôle de C.B.S. Scientific. C.B.S. Scientific ne pourr a en aucun cas être tenue responsable de dommages indirects, y compris, de m anière non limitative, la perte de bénéfices, le manque à gagner, la perte d'occasions d'aff aires, l'impos sibilité d'usage ou tous autres dommages associés, quelle qu'en soit la cause, ni de dom mages résultant de l'usage incorrect du produit.
DEUTSCH WICHTIGE INFORMATION FÜR DEN BENUTZER
Diese Bedienungsanleitung beschreibt wie man dieses Produkt sicher und wirksam benutzt. Bitte lesen und befolgen Sie alle Anweisungen in dieser Anleitung.
Das Dreieck mit Ausrufezeichen weist den Benutzer des Produktes darauf hin, daß wichtige Bedienungs-, Wartungs- und/oder Garantievorschriften zu beachten sind.
Das Dreieck mit Zickzackblitz warnt den Benutzer des Produktes vor möglichen Gefahren durch elektrische Spannungen.
Nichtbeachtung dieser Anweisungen kann zu persönlichen und/oder labortechnisc hen Schäden führen und gleichzeitig alle Garantien als nichtig erklären. Die DC Stromzufuhr muß imm er, vor dem Entfernen der Stromkabel vom Produkt, abges chaltet werden. Die Stromzufuhrkabel müssen zuerst von der Steckdose und erst dann vom Produkt entfernt werden. Um höchste Sicherheit zu gewährleisten sollte dieses System in einem abgesonderten und besonders ruhigen Ber eich eingesetzt werden und vor Unbefugten sicher sein.
GARANTIE UND HAFTUNG
Dieses Produkt wurde unter Anwendung von Produkten mit höchster Qualität und aus Materialien m it bester Verarbeitung und modernstem Des ign hergestellt. C.B.S Scientific gar antiert, daß das Produk t getestet wurde und alle publizierten Spezifikationen übertrifft. Diese Garantie ist jedoch nur gültig, wenn das Produkt nach der beigefügten Bedienungsanleitung bedient und gewartet wurde.
C.B.S. Scientific garantiert, daß dieses Produkt bei normaler Bedienung aus fehlerfreiem Material besteht und fehlerfrei in der Ausführung ist. Diese Garantie gilt für ein Jahr ab Lieferdatum. Sollte das Produkt in diesem Zeiraum fehlerhaft werden, bietet C.B.S. Scientif ic eine kostenlose Reparatur bzw. kostenlosen Ersatz, einschließlich freiem Rückporto. Diese Garantie schließt folgendes aus: Transportschaden, Schaden durch Nachlässigkeit, Mißbrauch oder Vernachlässigung, normale Abnützung durch regelmäßigen Gebrauch, Schaden durch Säureangriff, Schaden durch falsche Handhabung, Veränderung des Produktes durch den Benutzer, oder unzureichende Leistungen die sich nicht im Verantwortungsbereich von C.B.S. Scientific befinden. C.B.S. Sc ientific kommt unter keinen Umständen für folgende Schäden auf: Sachschadensverlust, Einkommensverlust, Verlust von Geschäftsmöglichkeiten, Verlust der Anwendung und andere damit verbundene Schäden die auf irgend eine Art und Weise entstanden sind, oder Schäden die aus falscher Anwendung des Produktes entstanden sind.
SECTION 1
ESPAÑOL INFORMACIÓN IMPORTANTE PARA EL USUARIO
El presente instructivo explica la manera de usar este producto en forma segura y efectiva. Sírvase leerlo en su totalidad y seguir detenidamente las indicaciones que contiene.
El símbolo del triángulo con exclamación llama la atención del usuario a requisitos importantes para el uso y mantenimiento del producto, así como para la validez de la garantía.
El símbolo del triángulo con rayo llama la atención del usuario a la posibilidad de riesgos eléctricos.
El incumplimiento de las instrucciones aquí señaladas podría dar lugar a riesgos a la persona, al laboratorio o a ambos y podría anular toda garantía. Siempre apague la fuente de corriente continua antes de desenchufar los cables eléctric os del producto. Primero desc onecte los cables de la fuente de energía y después del producto. Para mayor seguridad, siempre use este sistem a en un área aislada, de poco movimiento de personas e inaccesible a personal no autorizado. Jamás use equipo que presenta algún daño o fuga.
GARANTÍA Y RESPONSABILIDAD
Este producto fue fabricado de acuerdo con las normas más estrictas que sean factibles en cuanto a materiales, mano de obra y diseño. C.B.S Scientific garantiza que se sometió el producto a pruebas y que cumplirá o excederá las especif icaciones publicadas. Esta garantía s erá válida únicamente si se usa y se da servicio de mantenimiento al producto de acuerdo con las instrucciones señaladas.
C.B.S. Scientific garantiza que este producto se encontrará libre de defectos de m ateriales y mano de obra por un período de servicio normal de un año a partir de la fecha de embar que. Si el producto resulta defectuoso durante este período, C.B.S. Scientific lo reparará o lo repondrá, a criterio de C.B.S., libre de cargos, si se devuelve el producto a C.B.S. porte pagado. Esta garantía no cubre daños sufridos en tránsito, daños provocados por descuido, mal uso o negligencia, desgaste norm al como consecuencia del uso excesivo, daños atribuibles a corrosión provocada por solventes, daños causados por el uso indebido o alteraciones realizadas por el usuario ni rendimiento insatisfac torio atribuible a circunstancias fuera del control de C.B.S. Scientific. C.B.S. Scientific en ningún caso asumirá responsabilidad por daños incidentales o subsecuentes, inc luyendo, en forma no limitativa, la pérdida de utilidades, de ingresos, de oportunidades comerciales o del uso del pr oducto y otros daños afines, fuere cual fuere su origen, ni por daños derivados del uso incorrecto del producto.
ITALIANO INFORMAZIONI IMPORTANTI PER L’UTENTE
Questo manuale spiega come utilizzare questo prodotto in maniera sicura ed efficiente. Si pregai di leggere e seguire con cautela le istruzioni di ogni parte di questo manuale.
Il triangolo contenete il simbolo di un punto esclamativo avverte l’utente di importanti requisiti relativi al funzionamento, manutenzione e/o garanzia del prodotto.
Il triangolo contenete il simbolo di un lampo avverte l’utente del prodotto della
laboratorio, oltre a poter annullare la garanzia. Prima di distaccare il cordone d’alimentazione dal prodotto, spegnere sempre la sorgente di corrente continua. Distacc are i cordoni d’alimentazione prima dal lato della sorgente di tensione e poi dal lato del prodotto. Per maggior sicurezza, mettere sempre in funzione il prodotto in un’area isolata con poco traffico che non s ia acces sibile al personale non autorizzato. Non mettere mai in funzione un’apparecchiatura che sia danneggiata o abbia perdite.
GARANZIA E RESPONSABILITÀ
Questo prodotto è stato fabbricato seguendo gli s tandar d più elevati per i materiali, la manodopera e la progettazione. La C.B.S. Scientific garantisce il prodotto è stato sottopos to a prova e raggiunge o supera i valori pubblicati per i dati tecnici. Questa garanzia è valida solo se il prodotto è messo in esercizio e soggetto a manutenzione secondo le istruzioni fornite.
La C.B.S. Scientific garantisce che questo prodotto è liber o di difetti di materiali e manodopera, in normali condizioni d’esercizio, per la durata di un anno dalla data di spedizione. Se, in questo periodo, il prodotto si dimostrerà difettoso, la C.B.S. Scientific, a suo giudizio, lo riparerà o sostituirà. Questa garanzia non copre danni in transito, danni causati da negligenza, uso improprio, trascuratezza, normale consumo derivante da uso frequente, o danni causati da s olventi corrosivi, danni causati da maltrattamento o da modifiche apportate dall’utente e non copre pres tazioni insoddisfacenti c he siano il risultato di condizioni al di fuori del controllo del fabbricante. La C.B.S. Scientific non sarà in ogni caso responsabile per danni incidentali o consequenziali, incluso, senza limitazioni, perdite di profitto, perdita di entrate, perdita di opportunità d’affari e altri danni relativi, comunque causati, e per danni risultati da uso incorretto del prodotto.
possibilità di pericoli dovuti a corrente elettrica.
La mancata osservanza delle istruzioni può essere causa di pericolo alla propria persona ed al
C.B.S. Scientific 4 Dedicated Height Sequencer
General Information
1.1 Introduction
The C.B.S. Scientific Dedicated Height Nucleic Acid Sequencers feature safety covers with attached power leads, removable upper and lower reservoirs, epoxy-coated anodized aluminum heat dispersion plates, buffer drainage ports and a clear acrylic bar clamp. The Dual Dedicated Height Nucleic Acid Sequencers include the same features, with the convenience of divided upper reservoirs, if independent runs are required. The Sequencers are available in either a single or dual format and in 20cm or 33cm widths. Applications include Mutation Detection, Genotyping, Differential Display, DNA Footprinting, SDS Page, Microsatelites, Rnase Protection and Sequencing.
1.2 Specifications
Constructions: Upper buffer chamber Acrylic
Lower buffer chamber/base Acrylic Safety covers Acrylic Electrodes Platinum wire .012" diameter
Upright/base Acrylic Rotating base Acrylic
DC Power leads FR Urethane rated 7500VDC@ 200mA, 65ºC Bar Clamp Delrin and acrylic Heat dispersion plate Epoxy coated anodized aluminum
Glass plates Soda-lime float glass or
Low fluorescence glass Combs Polycarbonate, Mylar or White Spacers Polycarbonate, Mylar or White White spring clamps Polypropylene
Overall size
Nominal Unit Width 20cm unit 33cm unit
Single unit footprint: Overall width 30.5cm 41.0cm Overall height 32cm gel size 41.0cm 43.0cm 42cm gel size 51.0cm 53.0cm 52cm gel size 61.0cm 63.0cm 62cm gel size 71.0cm 73.0cm Dual unit footprint: Overall width 30.5cm 41.0cm Overall height 32cm gel size 41.0cm 43.0cm 42cm gel size 51.0cm 53.0cm 52cm gel size 61.0cm 63.0cm 62cm gel size 71.0cm 73.0cm
Recommended nominal buffer volumes Single units
upper reservoir 250ml 1500ml
lower reservoir Dual units with Divided Upper Reservoirs
upper reservoir lower reservoir
(per side) 300-500ml 400-600ml
(total volume/per side) 750/375ml 1500/750ml
(per side) 300-500ml 400-600ml
C.B.S. Scientific 5 Dedicated Height Sequencer
1.3 Safety
Power to the Sequencing unit must be supplied by an external DC voltage power supply that must be ground isolated so that the DC voltage output floats with respect to ground.
Current to the unit, provided from the external power supply, must enter the unit through the lid assembly, providing a safety interlock to the user. Current to the unit is broken when the lid is removed. Do not attempt to use the unit without the safety lid, and always turn the power
supply off before removing the lid, or when working with the unit in any way. Follow safety precautions specified by the power supply manufacturer.
C.B.S. Scientific 6 Dedicated Height Sequencer
C.B.S. Scientific 7 Dedicated Height Sequencer
Section 2 Description of Parts
2.1 Unpacking- Dedicated Height Sequencer
Please verify that your sequencing unit comes complete with the following components:
Buffer reservoirs, upper & lower with Leveling base with leveling feet safety covers with attached power leads Bar Clamp
Drain plugs installed in upper reservoir 2 white nylon tubing adapters for buffer
4 white spring clamps drain ports (Cat.# GPC-0001) Epoxy-coated anodized aluminum heat dispersion
4 knurled black thumb screws plate for securing lower reservoir to base reservoirs
If a kit was ordered verify that you received the following accessories:
1 comb (sharktooth or rectangular) 1 set spacers
1 set glass plates 1 roll gel sealing tape, cat. # GT-72-10
2 wedge plate separators
cat. # WPS-100 (not shown)
2.2 Unpacking- Dual Dedicated Height Sequencer
Please verify that your sequencing unit comes complete with the following components:
Buffer reservoirs, divided upper Rotating base with leveling feet
& lower with safety covers/power leads Bar Clamps (two) Drain plugs installed in upper reservoir Four white nylon tubing adapters for buffer Eight white spring clamps drain ports (Cat.# GPC-0001) Epoxy-coated anodized aluminum heat dispersion Six knurled black thumb screws plates (two) for securing lower reservoir to base reservoirs
If a kit was ordered verify that you received the following accessories:
Two combs (sharktooth or rectangular) Two sets spacers Two sets glass plates One roll gel sealing tape, cat. # GT-72-10 Two wedge plate separators Two temperature strip, cat.# EGT-100 (not shown)
cat. # WPS-100 (not shown)
Section 3
C.B.S. Scientific 8 Dedicated Height Sequencer
Assembly
3.1 Unit Assembly for Single Dedicated Height Sequencer
Place the Sequencer in a laboratory workspace for authorized personnel only. For protection against possible damage, the aluminum plate and upper and lower reservoirs are shipped detached from the unit. Do not attach reservoirs until after aluminum plate is installed. To assemble follow steps below:
A. Aluminum plate installation (see diagram below).
1. The top-front portion of the aluminum plate will be identified. Place the aluminum plate face down on the edge of the bench-top as illustrated below. Larger offset of screw holes in aluminum plate should align with the top of the sequencer upright.
2. Place the four white washers on top of screw holes.
3. Place sequencer in horizontal position with base overhanging bench-top making sure to support base of unit with one hand and line up the thumb screw holes with washer and aluminum plate holes.
4. Support base of unit with one hand while attaching the 4 thumbscrews. Tighten screws and return sequencer to upright position.
C.B.S. Scientific 9 Dedicated Height Sequencer
3.1 Unit Assembly for Single Dedicated Height Sequencer-continued
B. Installation of reservoirs.
1. Attach lower reservoir (with red power lead) by loosening black thumb screws on base of unit and sliding receptacle slots on reservoir around screw stem. Hand tighten.
2. Install upper reservoir (with black lead) by detaching upper black thumb-screws and positioning oval slot on upper reservoir so that they line up with screw holes on the base. White O-ring should be facing forward. Re-thread black screws through base of unit and hand tighten allowing reservoir to slide freely on sequencer upright. The upper reservoir remains free facilitating the clamping of the glass plate assembly to the reservoir using the bar clamp. See Section 4.1.
3. Upper and lower reservoirs are shipped with drain plugs installed. Verify that the drain plugs are secure. If you plan to use accessory Quick-Disconnect fittings (cat.# QDC-B and QDC-I), or ball valve, remove the drain plugs. Wrap the end of the Quick Disconnect Insert or valve with Teflon® tape and thread into the drain hole. Be sure the Quick Disconnect Body is secure and the Quick Disconnect Insert snaps into place.
4. Level the assembled sequencer using the four leveling feet supplied. Each sequencer is shipped with an attached bubble level.
3.2 Unit Assembly for Dual Dedicated Height Sequencer
Place the Sequencer in a laboratory workspace for authorized personnel only. To assemble follow steps below:
1. Attach lower reservoirs (with red power lead) by loosening black thumb screws on base of unit and sliding receptacle slots on reservoir around screw stem. Hand tighten.
2. Attach upper reservoir (with black leads) by detaching upper black thumb-screws and positioning oval receptacles on base of upper reservoir so that they line up with screw holes. White O-ring should be facing forward. Re-thread black screws through base of unit and hand tighten.
3. Upper and lower reservoirs are shipped with drain plugs installed. Verify that the drain plugs are secure. If you plan to use accessory Quick-Disconnect fittings (cat.# QDC-B and QDC-I), remove the drain plugs. Wrap the end of the Quick Disconnect Insert with Teflon tape and thread into the drain hole. Be sure the Quick Disconnect Body is secure and the Quick Disconnect Insert snaps into place.
4. Level the assembled sequencer using the four leveling feet supplied. Each sequencer is shipped with an attached bubble level.
C.B.S. Scientific 10 Dedicated Height Sequencer
3.3 Glass Plate Preparation
1. Select a pair of glass plates (if using a notched set, one notched and one unnotched back plate) for the appropriate gel height. Be sure the edges of the plates are nick or chip free.
2. It is vital that these plates are absolutely CLEAN ! Wearing gloves (and rinsing off any residual powder), clean the glass plates thoroughly with a non abrasive detergent and a plastic scouring pad. The cleaning solution should not leave a soap residue when rinsed thoroughly.
3. Rinse the glass plates thoroughly with de-ionized water and wipe dry with a lint free paper towel.
4. If desired, treat the inside (acrylamide contact) surface of one (usually the notched plate) or both glass plates with a silanizing reagent, such as C.B.S. Scientific's Gel Repel, following the manufacturer's instructions.
5. Immediately before assembling the glass plate 'sandwich', rinse the inside surfaces with 95% ethanol and wipe dry with a lint-free paper towel. Avoid touching the inside surfaces with your fingers. The wearing of gloves is highly recommended. Impeccably clean, dust-free plates are essential for pouring gels without bubbles.
6. Assemble the glass plate sandwich in the conventional manner; place the back glass plate inside-up on the bench, align the cleaned spacers at the sides, and place the notched plate inside-down on the side spacers. **The use of metal “bulldog” clamps is not recommended.
C.B.S. Scientific 11 Dedicated Height Sequencer
#GPC
000
3.4 Methods for sealing glass plate sandwich
(A) Gel Slider, ( B) Capillary Gel Casting or (C) Yellow tape
(A) C.B.S. Scientific's Gel Slider, Sequencing Gel Caster. This adjustable width unit
simplifies the pouring of problem-free sequencing gels for the novice or experienced sequencer. Unit sold separately, see instructions for Gel Slider and skip section 3.5.
(B) Capillary Gel Casting. Depending upon user preference, capillary gel casting can be
performed (1) by using a bottom spacer at the bottom of the gel plate assembly, (2) by taping the bottom section with yellow tape, or (3) by leaving the end completely open. Assemble glass plate sandwich with side spacers on a level working surface.
1. If using a bottom spacer, offset the side spacers to accommodate the bottom spacer, so the spacers line up with the edges of the back plate. Clamp continuously with casting clamps (cat.# GPC-0002) as shown in illustration.
2. If using tape for the bottom section of the glass plate assembly, extend the tape around the bottom corners in both directions to ensure an adequate seal. Apply the tape as smoothly as possible to avoid forming air channels or bubbles along the edges of the glass plates. Rub the tape firmly onto glass to eliminate air channels and ensure a liquid-tight seal. Clamp the side sections of the glass plate assembly with casting clamps (cat.# GPC-0002).
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2
3. If leaving the bottom of the glass plate assembly open, clamp the side sections of the glass plate assembly with casting clamps (cat.# GPC-0002).
C.B.S. Scientific 12 Dedicated Height Sequencer
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