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  • 1

    COMPAC-50 Comet Assay System

    Von Cleaver Scientific

    9963.1

    Komplettsystem zur schnellen Einzelzell-Gelelektrophorese mit hohen Durchsatzraten

    Platzsparendes, kompaktes System

    Kammer und Zubehr zum Schutz der DNA-Proben aus schwarzem, lichtundurchlssigem Acryl

    2 Objekttrgerhalter zur Bearbeitung von bis zu 50 Objekttrgern gleichzeitig

    Zeitsparende Elektrophorese (Dauer pro Lauf ca. 20 min)

    Integrierte Keramik-Khlplatte mit Khlakku, kein zustzlicher Umlaufkhler ntig

    1. PACKLISTE

    Anz. Artikel Bezeichnung 1 Comet Assay Tank (Kammer mit Sicherheitsdeckel + integrierten Stromanschlssen, 4 mm Stecker) 1 Paar Elektrophoresekabel 2 Objekttrgerhalter (fr je 25 Objekttrger) 10 Behlter mit Deckel, passend fr Objekttrgerhalter 1 Khlakku 1 Libelle zur Nivellierung

    2. ZUBEHR

    Objekttrgerhalter (fr 25 Objekttrger) (1 Stck) 9964.1 Behlter fr Objekttrgerhalter (1 Stck) 9965.1 Behlter fr Objekttrgerhalter (4 Stck) 9984.1 Plus-Elektrode (1 Stck) 9966.1 Minus-Elektrode (1 Stck) 9967.1 Elektrophoresekabel Rotiphorese

    PROfessional ( 1 Paar ) 6848.1

    3. TECHNISCHE DATEN

    Abmessungen (B x T x H) 26,5 x 15 x 15 cm

    Beladungskapazitt Gert insg. 50 Objekttrger (25 x 75 mm)

    Beladungskapazitt pro Halter 25 Objekttrger (25 x 75 mm)

    Volumen 550 ml

    Empfohlene Laufbedingungen 27 V / 450 mA fr 20 min

    Empfohlene Power Supplies Roth-STANDARD, EV2310, EV2650

    BEWAHREN SIE BITTE DIE VERPACKUNG BIS ZUM ABLAUF DER GARANTIEZEIT AUF.

    Weitere Informationen zum Produkt finden Sie in der anhngenden Cleaver Gebrauchsanleitung.

    COMPAC-50 Comet Assay System 9963.1

  • 2

    Carl Roth GmbH+Co. KG

    Schoemperlenstrae 3-5 76185 Karlsruhe Postfach 100121 76231 Karlsruhe Tel.: (+49)721/5606-0 Fax: (+49)721/5606-149 E-Mail: info@carlroth.de Internet: www.carlroth.de e.d. 12/2016

    COMPAC-50 Comet Assay System

    By Cleaver Scientific

    9963.1 Complete system for rapid high throughput single cell gel electrophoresis (SCGE)

    Space-saving, compact system Chamber and accessories are made of ebony acrylic material to protect DNA samples against

    light 2 slide carriers for processing of up to 50 slides simultaneously Time-saving electrophoresis (length per run ca. 20 min) Integrated ceramic cooling platform with cool pack, no additional chiller necessary

    1. PACKING LIST

    No. Items Description 1 Comet Assay Tank (Chamber with safety lid + integral power leads, 4 mm plugs) 1 Paar Elektrophoresis Cables 2 Slide Carrier (for 25 slides each) 10 Treatment Dishes, suitable to slide carrier 1 Cool Pack 1 Level Bubble

    2. ACCESSORIES

    Slide Carrier (for 25 slides) (1 carrier) 9964.1 Treatment Dish for slide carrier (1 dish) 9965.1 Treatment Dish for slide carrier (4 dishes) 9984.1 Positive Electrode (1 electr.) 9966.1 Negative Electrode (1 electr.) 9967.1 Electrophoresis Cable Rotiphorese

    PROfessional (1 pair) 6848.1

    3. TECHNICAL DATA

    Dimensions (W x L x H) 26,5 x 15 x 15 cm

    Total Slide Capacity 50 slides (25 x 75 mm)

    Slide Capacity per Rack 25 slides (25 x 75 mm)

    Volume 550 ml

    Recommended Running Conditions 27 V / 450 mA for 20 min

    Recommended Power Supplies Roth-STANDARD, EV2310, EV2650

    PLEASE RETAIN ALL PACKAGING MATERIALS UNTIL THE WARRANTY PERIOD HAS EXPIRED.

    For further Information see Cleaver Instruction

    Manual below.

    COMPAC-50 Comet Assay System

    9963.1

    mailto:info@carlroth.dehttp://www.carl-roth.de/

  • 3

    Instruction Manual

    COMPAC-50 High Throughput Comet Electrophoresis System

  • 4

    Table of content

    Page

    Section 1 Safety Information

    1.1 General safety Instructions 1.2 General Laboratory Safety

    5

    6

    Section 2 General Information

    2.1 Introduction

    2.2 Product Information 2.3 Packing List 2.4 Specifications 2.5 Care and Maintenance

    7

    9

    10

    10

    11

    Section 3 Operating Instructions

    3.1 Equipment and Materials to be Supplied by User 3.2 Reagents

    3.3 Operation of Equipment

    3.4 Optimization of running conditions 3.5 Related products from Cleaver Scientific 3.6 Cleaver Scientific Chilling Plate

    12

    13

    13

    14

    15

    16

    Section 4 Comet Assay Protocols

    17

    Section 5 Warranty

    21

  • 5

    Section 1 Safety Information

    1.1) General Safety Instructions

    WHEN USED CORRECTLY, THESE UNITS POSE NO HEALTH RISK. HOWEVER, THESE

    UNITS CAN DELIVER DANGEROUS LEVELS OF ELECTRICITY AND ARE TO BE OPERATED

    ONLY BY QUALIFIED PERSONNEL FOLLOWING THE GUIDELINES LAID OUT IN THIS

    INSTRUCTION MANUAL.

    ANYONE INTENDING TO USE THIS EQUIPMENT SHOULD READ THE COMPLETE MANUAL

    THOROUGHLY.

    THE UNIT MUST NEVER BE USED WITHOUT THE SAFETY LID CORRECTLY IN POSITION.

    THE UNIT SHOULD NOT BE USED IF THERE IS ANY SIGN OF DAMAGE TO THE

    EXTERNAL TANK OR LID.

    THESE UNITS COMPLY WITH THE STATUTORY CE SAFETY DIRECTIVES: 73/23/EEC:

    LOW VOLTAGE DIRECTIVE: IEC 1010-1:1990 plus AMENDMENT 1:1992

    EN 61010-1:1993/BS EN 61010-1:1993

  • 6

    1.2) General laboratory safety

    Disposable gloves and a laboratory coat must be worn when carrying out the assay.

    Safety goggles must be worn when using hazardous chemicals and where there is a

    risk of reagents splashing.

    All cell samples must be treated as potentially hazardous. Avoid contact with skin

    and eyes.

    All appropriate COSHH forms must be read prior to starting the assay, and reagent

    disposal requirements must be adhered to. Care must be taken when handling

    hydrochloric acid, sodium hydroxide and propidium iodide. Avoid contact with skin

    and eyes.

    Small amounts of cell waste can be disposed of via autoclave waste. To large

    amounts of cell culture waste, add Presept disinfectant tablet, leave overnight and

    dispose of down the sink following day.

    Glass coverslips and slides must be disposed of in sharps bins, for incineration.

    Do not pour agarose down the drains. Agarose must be disposed of via autoclave

    waste.

    Precautions must be taken when heating reagents in the microwave. Use heat proof

    gloves.

    Precautions must be taken when handling the cells. In case of spillage, clean with

    70% IMS (Industrial methylated spirit).

  • 7

    Section 2 General Information

    2.1) Introduction

    The comet assay is a process by which DNA damage (strand breaks) may be

    quantified in individual cells. Under alkali conditions, other DNA damage products, known as alkali-labile sites may also be detected. Furthermore, the use of DNA repair enzymes may provide a greater degree of specificity by revealing DNA

    nucleobase damage. An overview of the assay is:

    1) A single cell suspension of the cells under investigation is mixed with low

    melting point agarose.

    2) The cell/agarose mix is layered on glass microscope slides, pre-coated with

    agarose, and the agarose allowed to set.

    3) The cells are lysed under high pH.

    4) The presence of strand breaks and high pH allows the cellular DNA to

    unwind. Electrophoresis draws the DNA out of the nucleoid body forming a

    tail. The amount of migration (the amount of DNA in the tail versus the

    head) is proportional to the initial amount of DNA damage.

    5) The high pH is neutralised by neutralization buffer and washing the slides.

    6) The DNA is stained and then again washed.

    7) The Comets are visualised and scored for DNA damage assessment, using

    fluorescent microscopy and image analysis software.

    This process is laborious and time-consuming, having to treat every slide individually, in particular through steps 3-6. Placing the microscope slides flat, on

    their largest face during electrophoresis (step 4) limits the number of slides that can fit in the electrophoresis tank.

    Figure 1 COMPAC-50 High Throughput Comet Electrophoresis System

  • 8

    Cleaver Scientifics COMPAC-50 High Throughput Comet Electrophoresis System

    provides a simple and effective method to process a greater number of slides simultaneously through steps 3-6 by placing the slides on their longest edge in a

    microscope slide carrier (Figure 2).

    Figure 2 COMPAC-50 microscope slide carrier

    Coplin jars, in which lysis is performed normally in conventional comet assay carry

    a maximum of 10 slides. Microscope slide carriers however can accommodate 25 slides on their side, and can be used in conjunction with treatment dishes to perform lysis, neutralisation and staining steps (steps 3,5 and 6), together with all

    the associated washing steps (Figure 3). Manipulating 25 slides at a time would vastly increase the speed of the process. Also changing the conventional style of

    electrophoresis tank for comet assay, to one which could accommodate the depth of the microscope slides carrier would mean slides could be transferred to the electrophoresis tank and electrophoresed whilst still in the slide carrier.

    Furthermore, through using COMPAC-50 High Throughput Comet Electrophoresis System, the number of slides that can be electrophoresed is increased in a considerably smaller footprint as a standard comet tank. Decreasing the handling

    of the slides will decrease the risk of possible damage or loss of LMP gels from the microscope slides,

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