C.B.S. Scientific SG User Manual

INSTRUCTION MANUAL

Adjustable Height Nucleic Acid

Sequencing Systems

Single Units Dual Units

SG-400-20 DASG-400-20
SG-500-20 DASG-500-20
SG-600-20 DASG-600-20
SG-800-20
SG-1000-20
SG-400-33 DASG-400-33
SG-500-33 DASG-500-33
SG-600-33 DASG-600-33
SG-800-33
SG-1000-33

TABLE OF CONTENTS

Page

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-Adjustable Height Sequencer 7

2.2 Unpacking-Dual Adjustable Height Sequencer 8

Section 3 Assembly

3.1 Unit assembly for Single Adjustable Sequencer 9

3.2 Unit assembly for Dual Adjustable Sequencer 9

3.3 Glass Plate Preparation 10

3.4 Methods for Sealing Glass Plate Sandwich 11-12

3.5 Pouring the Gel 13-14

Section 4 Electrophoresis

4.1 Pre-Electrophoresis 15-16

4.2 Loading Samples 16

4.3 Electrophoresis 17

4.4 Post Electrophoresis-Gel Processing 18-19

4.5 Recommended Buffers and Reagents 20

4.6 References and Suggested Reference Literature 21

Section 5 Tables: Power Settings, Troubleshooting, Comb Volumes

5.1 Recommended Power Settings 22

5.2 Troubleshooting Guide 23

5.3 Comb Volumes 24-26

Section 6 Maintenance of Equipment

6.1 Care and Handling 27

6.2 Maintenance 27

Section 7 Ordering Information 28-36

C.B.S.  Scientific 2 Adjustable Height Sequencer

ENGLI

S

H

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 electrical

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.

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 Adjustable Height Sequencer

FRANÇAIS INFORMATION IMPORTANTE À L'USAGE DES UTILISATEURS

Le présent manuel d'utilisation explique la manière de se ser vir efficacement du produit en c onditions

de sécurité. Il est recommandé de soigneusem ent 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.

ESPAÑOL INFORMACIÓN IMPORTANTE PARA EL USUARIO

El presente instructivo explica la manera de usar este producto en f orma segura y efectiva. Sírvase
leerlo en su totalidad y seguir detenidamente las indicaciones que contiene.

El símbolo del triángulo con exclamación llama la atenc ión 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 som etió el produc to 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.

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 Zickzack blitz 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 Design herges tellt. C.B.S. Scientif ic garantiert, 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.

ITALIANO INFORMAZIONI IMPORTANTI PER L’UTENTE

Questo manuale spiega come utilizzare questo prodotto in maniera sicura ed eff iciente. 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 impor tanti
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 Adjustable Height Sequencer

SECTION 1
General Information

1.1 Introduction

The C.B.S. Scientific Adjustable Nucleic Acid Sequencers features the flexibility of running path
lengths from 22cm up to 1 meter. This adjustable feature allows for rapidly changing research
priorities for the individual laboratory researcher. More recent state-of-the-art techniques have
been adapted for use on our adjustable units including large scale screening for Gel Mobility
Shifts (gel retardation-MSA), mRNA Differential Display using RT-PCR, DNA footprinting, RNase
protection assays, as well as the classical methods of nucleic acid sequencing either by Sanger­Coulson (dideoxy method) or the original Maxam-Gilbert reaction procedure. The units are
available in either 20cm or 33cm wide formats.

1.2 Specifications
Constructions:

Upper buffer chamber Acrylic

Lower buffer chamber/base Acrylic
Safety covers Acrylic
Electrodes Platinum wire .012" diameter
DC Power leads FR Urethane rated 7500VDC
200mA, 65ºC
Guide rods Delrin
Heat dispersion plate Anodized aluminum
Glass plates Soda-lime float glass or Low fluorescent
glass
Combs Polycarbonate, Mylar or White
Spacers Polycarbonate, Mylar or White
White spring clamps Polypropylene

Overall size
Footprint
Base size-20cm unit 30.5cm (l) x 30.5cm (w)
Base size-33cm unit 40.6cm (l) x 30.5cm (w)
Height 42, 52, 62, 82, 102cm

Recommended buffer volumes

20cm wide units
upper reservoir 700ml

lower reservoir 700ml
33cm wide units
upper reservoir 33cm 1400-1600ml
lower reservoir 33cm 1000-1600ml

C.B.S.  Scientific 5 Adjustable Height Sequencer

1.3 Safety

Power to the Sequencing unit is to 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 alw ays turn the power

supply off before removing the lid, or when working w ith the unit in any way. Follow
safety precautions specified by the power supply manufacturer.

C.B.S.  Scientific 6 Adjustable Height Sequencer

C.B.S.  Scientific 7 Adjustable Height Sequencer

Section 2
Description of Parts

2.1 Unpacking- Adjustable 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
Drain plugs installed in upper reservoir • Guide Rods

• 6 white spring clamps • 2 white nylon tubing adapters for buffer
(Cat.# GPC-0001) drain ports

• 2 knurled black thumb screws • Rosette knobs (three)
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 • 1 temperature strip, Cat.# EGT-100 (not shown)

Cat. # WPS-100 (not shown)

2.2 Unpacking- Dual Adjustable 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  Rosette knobs (two)
 Drain plugs installed in upper reservoir  Four white nylon tubing adapters for buffer
 Twelve white spring clamps drain ports
(Cat.# GPC-0001)  Guide Rods
 Four knurled black thumb screws
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)

C.B.S.  Scientific 8 Adjustable Height Sequencer

Section 3
Assembly

3.1 Unit Assembly for Adjustable Height Sequencer

1. Place the Adjustable Sequencer on a level work surface in an authorized work area. Attach

three guide rods to base. Tilt base on its side, place stainless steel screw through
countersunk hole and screw the guide rods onto the base.

2. Secure lower reservoir to base by tightening the two black knurled thumb screws.

3. Once guide rods are secure and apparatus is flat on laboratory bench, slide the upper

reservoir over the three guide rods. To adjust upper reservoir to desired height, align notch
in glass plate with upper reservoir fascia. Tighten the three rosette knobs on the upper
reservoir until secure. Level apparatus using leveling feet in base.

4. 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.

5. 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 Adjustable Height Sequencer

1. Place the Dual Adjustable Sequencer on a level work surface in an authorized work area.

Attach two guide rods to base. Tilt base on its side, place stainless steel screw through
countersunk hole and screw the guide rods onto the base.

2. Secure lower reservoirs to base by tightening the four black knurled thumb screws.

3. Once guide rods are secure and apparatus is flat on laboratory bench, slide the divided

upper reservoir over the two guide rods. To adjust upper reservoir to desired height, align
notch in glass plate with upper reservoir fascia. Tighten the two rosette knobs on the upper
reservoir until secure. Level apparatus using leveling feet in base.

4. 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.

5. Dual units include a rotating base. Align cut-out in bottom of base with rotating base. Use

stationary pin to keep unit form rotating.

6. Level the assembled sequencer using the four leveling feet supplied. Each sequencer is

shipped with an attached bubble level.

C.B.S.  Scientific 9 Adjustable 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.

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.

C.B.S.  Scientific 10 Adjustable Height Sequencer

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).

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).

GPC-0002

C.B.S.  Scientific 11 Adjustable Height Sequencer