VTi 50 ROTOR
Contents
1. Volume mL 7 td 20 10 0 a 39 mL tube 27 25 20 15 10 5 O lt 27 mL tube 19 be tt hte 15 10 5 O lt 15 mL tube 1 8 173 9 f 40 000 854 Le 48 F rpm Se 1 7 30 00 p R Tiel 40 poe f AY wi ei on goo pos 1 6 re 6 S lt p J OY KN E er J oN 5 1 5 DSF 5 oA S gt a 1 4 VTi 50 ROTOR 20 C 1 3 4 C 1 2 1 1 02 4 6 8 10 12 14 16 18 20 22 2425 Distance Across Tube Diameter mm During Centrifugation Figure 2b CsCl Gradients at Equilibrium for the VTi 50 Rotor Centrifugation of homogeneous CsCl solutions at the maximum allowable speeds from Figure 2a results in gradients presented here 15 VTi 50 Rotor AT EQUILIBRIUM During or After Centrifugation2. Volume mL 39 30 20 10 O 39 mL tube l 27 25 20 15 10 5 O lt 27 mL tube 1 9 15 10 5 0 15 mL tube or 20 C Lb 1 8 4 207 only these a Wwolour eso z SSSNNREP me pz Pai L aT ya ltt Ge 1 7 j Zz Pa Cgi L7 Pani Bai cad 4 ral A y LA Lier eal Jr 1 6 Trt l ars et a 1 J Lt F AT ee L ET pa 15 9 ma 1 5 z z rai 4 Ea ie 2 Att gt Ga zai 3B P7 iat a amp Lert AAG 1 4 pra Tt Cai T peT A L Lt a Pal 1 3 VTi 50 ROTOR 50000 12 40000 30 000 1 1 02 4 6 8 10 12 14 16 18 20 22 2425 Distance Across Tube Diameter mm During Centrifugation Figure 2c CsCl Gradients for Lower Densities Densities used to generate curves are printed along the curves 16 VTi 50 Rotor between bands will be greatest The gradients in Figures 2b and 2c can be generated from step or linear gradients or from homogeneous solutions But the total amount of CsCl in solution must be equivalent to a homogeneous solution corresponding to the concentrations specified TYPICAL EXAMPLES FOR DETERMINING CsCl RUN PARAMETERS Example A Knowing homogeneous CsCl solution density 1 71 g mL and approximate particle buoyant densities 1 70 and 1 65 g mL at 20 C where will particles band 1 In Figure 2a find the curve that corresponds to the required run temperature 20 C The maximu
3. Ontario Canada L5N 7Y2 Telephone 905 819 1234 Fax 905 819 1485 CHINA Beckman Coulter Inc Beijing Representative Office Unit 2005A 2006 2009 East Ocean Center Jian Guomenwai Avenue Beijing 100004 China Telephone 86 10 6515 6028 Fax 86 10 6515 6025 6515 6026 EASTERN EUROPE MIDDLE EAST AFRICA Beckman Coulter International S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 41 22 994 07 07 Fax 41 22 994 07 00 FRANCE Beckman Coulter France S A Paris Nord II 33 66 rue des Vanesses B P 50359 Villepinte France 95942 ROISSY CDG Cedex Telephone 01 49 90 90 00 Fax 01 49 90 90 10 e mail bioresearch france beckman com GERMANY Beckman Coulter GmbH Siemensstrasse 1 D 85716 Unterschleissheim Lohhof Germany Telephone 89 35870 0 Fax 89 35870 490 e mail bioresearch de beckman com rroo COULTER Beckman Coulter Inc HONG KONG Beckman Coulter Hong Kong Ltd 12th Floor Oxford House 979 King s Road Taikoo Place Hong Kong Telephone 852 2814 7431 2814 0481 Fax 852 2814 1599 2873 4511 ITALY Beckman Coulter S p a Centro Direzionale Lombardo Palazzo F 1 Via Roma 108 20060 Cassina de Pecchi Milano Italy Telephone 02 953921 Fax 02 95392264 e mail beckman_italia beckman com JAPAN Beckman Coulter K K Toranomon 37 Mori Bldg 3 5 1 Toranomon Minato ku Tokyo 105 0001 Japan Telephone 03 5
4. Rotor RUN TIMES TIME HR MIN The k factor of the rotor is a measure of the rotor s pelleting aa efficiency Beckman Coulter has calculated the k factors for all of Hi ju i j a j its preparative rotors at maximum rated speed and using full tubes The k factor is calculated from the formula ks In max min io X FENA 1 o2 3600 where is the angular velocity of the rotor in radians per second 0 105 x rpm Fmax is the maximum radius and rmin is the minimum radius After substitution 253x 10 D1n nax rnin A rpm Use the k factor in the following equation to estimate the run time t in hours required to pellet particles of known sedimentation coeffi cient s in Svedberg units S f 3 Run times can be estimated for centrifugation at less than maximum speed by adjusting the k factor as follows 2 50 000 4 kaaj aaa run speed Run times can also be estimated from data established in prior exper iments if the k factor of the previous rotor is known For any two rotors a and b t k o b b For more information on k factors see Use of k Factor for Estimating Run Times from Previously Established Run Conditions publication DS 719 10 VTi 50 Rotor SLOW ACCELERATION DECELERATION Vertical banding of sample and gradient formation occurs with centrifugation With deceleration tube contents reorient back to horizontal position For gradient stability when preformed gradients are u
5. Ultracentrifuge Rotors 1 year No Proration For Zonal Continuous Flow Component Test and Rock Core ultracentrifuge rotors see separate warranty Warranty Conditions as applicable 1 This warranty is valid for the time periods indicated above from the date of shipment to the original Buyer by Beckman Coulter or an authorized Beckman Coulter representative 2 This warranty extends only to the original Buyer and may not be assigned or extended to a third person without written consent of Beckman Coulter 3 This warranty covers the Beckman Coulter Centrifuge Systems only including but not limited to the centrifuge rotor and accessories and Beckman Coulter shall not be liable for damage to or loss of the user s sample non Beckman Coulter tubes adapters or other rotor contents 4 This warranty is void if the Beckman Coulter Centrifuge Sys tem is determined by Beckman Coulter to have been operated or maintained in a manner contrary to the instructions in the operator s manual s for the Beckman Coulter Centrifuge System components in use This includes but is not limited to operator misuse abuse or negligence regarding indicated main tenance procedures centrifuge and rotor classification require ments proper speed reduction for the high density of certain fluids tubes and tube caps speed reduction for precipitating gradient materials and speed reduction for high temperature operation 5 Rotor bucket
6. mL polyallomer pkg 56 aluminum spacer 242 000 x g 36 25 x 64mm Quick Seal 344323 aluminum spacer 342417 348124 50 000 rom 27 mL Ultra Clear pkg 50 242 000 x g floating spacer 343448 36 25x64mm Quick Seal 343665 aluminum spacer 342417 348124 50 000 rom 27 mL polyallomer pkg 50 242 000 x g floating spacer 343448 36 25x35mm Quick Seal 344324 aluminum spacer 342417 348124 50 000 rom 15 mL Ultra Clear pkg 50 242 000 x g floating spacer 343448 28 25x38mm Quick Seal 343664 aluminum spacer 342417 348124 50 000 rom 15 mL polyallomer pkg 50 242 000 x g floating spacer 343448 28 Includes disposable plastic plugs t Floating spacers part of the g Max system of tube support are made of Noryl a registered trademark of GE Plastics Use two floating spacers with this tube 25 C a gt Gold Aluminum a S Pug Aluminum Spacer ba Dome Pa shaped tube Metal Spacer SS Floating P Spacer Bell top 7 tube VTi 50 Rotor Temperature Limits e Plastic tubes have been centrifuge tested for use at temperatures between 2 and 25 C For centrifugation at other temperatures pretest tubes under anticipated run conditions e If tubes are frozen before use make sure that they are thawed to at least 2 C prior to centrifugation OptiSeal Tubes OptiSeal tubes come with plastic plugs and can be quickly and easily prepared for use without tools
7. of a cotton swab or similar nonmetallic tool to pry the gasket from the plug Do this carefully so that the plug is not damaged The new gasket snaps onto the grooved end of the plug Refer to Appendix A in Rotors and Tubes for the chemical resistances of rotor and accessory materials Your Beckman Coulter representa tive provides contact with the Field Rotor Inspection Program and the rotor repair center CLEANING Rotor Cleaning Kit 339558 Gasket Threads a r DECONTAMINATION VTi 50 Rotor Wash the rotor and rotor components immediately if salts or other corrosive materials are used or if spillage has occurred Do not allow corrosive materials to dry on the rotor Under normal use wash the rotor frequently at least weekly to prevent buildup of residues 1 Wash the rotor plugs and spacers in a mild detergent such as Beckman Solution 555 339555 that won t damage the rotor The Rotor Cleaning Kit 339558 contains two plastic coated brushes and two quarts of Solution 555 for use with rotors and accessories Dilute the detergent 10 to 1 with water gt NOTE Do not wash rotor components in a dishwasher Do not soak in detergent solution for long periods such as overnight 2 Rinse the cleaned rotor and components with distilled water 3 Air dry the rotor upside down Do not use acetone to dry the rotor Clean plug threads as necessary Use a brush and concentrated Solution 555 Rinse a
8. same density Make sure that cavities in use have the proper spacers inserted before installing the rotor plugs If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories if pathogenic or radioactive materials are involved Never exceed the maximum rated speed of the rotor and labware in use Refer to the section on RUN SPEEDS and derate the run speed as appropriate Do not use sharp tools on the rotor that could cause scratches in the rotor surface Corrosion begins in scratches and may open fissures in the rotor with continued use VTi 50 Rotor VTi 50 ROTOR min fav Tmax i U S Pat Nos 4 102 490 4 290 550 Axis of Ree Japanese Pat No 1 469 154 SPECIFICATIONS Maximum speed 0 0 00 e eee eee eee 50 000 rpm Density rating at maximum speed 00 1 7 g mL Relative Centrifugal Field at maximum speed At fnar 86 6 mm 2 2 ee eee 242 000 x g At ray 13 7 TMD shod hh bts he bed heed 206 000 x g At rnin 00 8MM cose kee eee eee nee 170 000 x g k factor at maximum speed 0 0 eee cece eee eee 36 Conditions requiring speed reductions see RUN SPEEDS Number of tube cavities 0 0 eee eee 8 Available tubes oo os os kee be chee eee sheet eae see Table 1 Nominal tube dimensio
9. sets purchased concurrently with or subsequent to the purchase of a Swinging Bucket Rotor are warranted only for a term co extensive with that of the rotor for which the bucket sets are purchased 6 This warranty does not cover the failure of a Beckman Coulter rotor in a centrifuge not of Beckman Coulter manufacture or if the rotor is used in a Beckman Coulter centrifuge that has been modified without the written permission of Beckman Coulter or is used with carriers buckets belts or other devices not of Beckman Coulter manufacture 7 Rotor parts subject to wear including but not limited to rotor O rings VTi NVT TLV MLN and TLN rotor tube cavity plugs and gaskets tubing tools optical overspeed disks bear ings seals and lubrication are excluded from this warranty and should be frequently inspected and replaced if they become worn or damaged 8 Keeping a rotor log is not mandatory but may be desirable for maintenance of good laboratory practices Repair and Replacement Policies 1 Ifa Beckman Coulter rotor is determined by Beckman Coulter to be defective Beckman Coulter will repair or replace it subject to the Warranty Conditions A replacement rotor will be warranted for the time remaining on the original rotor s warranty 2 Ifa Beckman Coulter centrifuge is damaged due to a failure of a rotor covered by this warranty Beckman Coulter will supply free of charge i all centrifuge parts required for repair ex
10. 404 8359 Fax 03 5404 8436 MEXICO Beckman Coulter de Mexico S A DE C V Avenida Popocat petl 396 Colonia Gral Pedro Maria Anaya Codigo Postal 03340 Maxico D F Mexico Telephone 525 605 77 70 Fax 525 575 18 25 NETHERLANDS Beckman Coulter Nederland B V Nijverheidsweg 21 3641 RP Mijdrecht Postbus 47 3640 AA Mijdrecht The Netherlands Telephone 0297 230630 Fax 0297 288082 SINGAPORE Beckman Coulter Singapore Pte Ltd 116 Changi Road Unit 03 01 02 Singapore 419718 Telephone 65 339 3633 Fax 65 336 6303 SOUTH AFRICA Beckman Coulter Stand 1A Primegro Park Tonetti Street 1685 Halfway House Johannesburg Republic of South Africa Telephone 27 11 805 2014 5 Fax 27 11 805 4120 SPAIN Beckman Coulter Espa a S A C Caleruega 81 28033 Madrid Spain Telephone 34 91 3836080 Fax 34 91 3836096 email bioresearch espana beckman com SWEDEN Beckman Coulter AB Archimedesvaegen 7 Box 111 56 SE 168 11 Bromma Sweden Telephone 0 8 564 85 900 Telefax 0 8 564 85 901 SWITZERLAND Beckman Coulter International S A 22 Rue Juste Olivier Case Postale 301 303 CH 1260 Nyon Switzerland Telephone 0800 850 810 Fax 0848 850 810 TAIWAN Beckman Coulter Taiwan Inc Taiwan Branch 8th Floor 216 Tun Hwa South Road Section 2 Taipei 106 Taiwan Republic of China Telephone 886 2 2378 3456 Fax 886 2 2377 0408 TURKEY Beckman Coulter Ltd E 5 Yanyol Faith C
11. CIAL OR CONSEQUENTIAL DAMAGES OF ANY KIND WHATSOEVER ARISING OUT OF THE MANUFAC TURE USE SALE HANDLING REPAIR MAINTENANCE OR REPLACEMENT OF THE PRODUCT Factory Rotor Inspection Service Beckman Coulter Inc will provide free mechanical and metallurgical inspection in Palo Alto California USA of any Beckman Coulter rotor at the request of the user Shipping charges to Beckman Coulter are the responsibility of the user Rotors will be inspected in the user s laboratory if the centrifuge in which they are used is covered by an appropriate Beckman Coulter Service Agreement Contact your local Beckman Coulter office for details of service coverage or cost Before shipping contact the nearest Beckman Coulter Sales and Service office and request a Returned Goods Authorization RGA form and packaging instructions Please include the complete rotor assembly with buckets lid handle tube cavity caps etc A SIGNED STATEMENT THAT THE ROTOR AND ACCESSO RIES ARE NON RADIOACTIVE NON PATHOGENIC NON TOXIC AND OTHERWISE SAFE TO SHIP AND HANDLE IS REQUIRED Beckman Coulter Worldwide Life Science Research Division Offices AUSTRALIA Beckman Coulter Australia Pty Ltd Unit D 24 College St Gladesville NSW 2111 Australia Telephone 61 2 9844 6000 or toll free 1 800 060 880 Fax 61 2 9844 6096 email lifescienceaustralia beckman com CANADA Beckman Coulter Canada Inc 6755 Mississauga Road Suite 600 Mississauga
12. ED L5 TB 050P ci BECKMAN ee COULTER VTi 50 ROTOR Used In Beckman Coulter Class H R and S Preparative Ultracentrifuges 2001 Beckman Coulter Inc PUBLISHED BY THE SPINCO BUSINESS CENTER OF BECKMAN COULTER INC PALO ALTO CALIFORNIA 94304 VTi 50 Rotor SAFETY NOTICE This safety notice summarizes information basic to the safe use of the rotor described in this manual The international symbol displayed above is a reminder to the user that all safety instructions should be read and understood before operation or maintenance of this equipment is attempted When you see the symbol on other pages throughout this publication pay special attention to the specific safety information presented Observance of safety precautions will also help to avoid actions that could damage or adversely affect the performance of the rotor This rotor was developed manufactured and tested for safety and reliability as part of a Beckman Coulter ultracentrifuge rotor system Its safety or reliability cannot be assured if used in a centrifuge not of Beckman Coulter s manufacture or in a Beckman Coulter ultracentrifuge that has been modified without Beckman Coulter s approval A SiS i ie Le Handle body fluids with care because they can transmit disease No known test offers complete assurance that such fluids are free of micro organisms Some of the most virulent Hepatitis B and C viruses HIV I V atypical mycobacteria and certain sy
13. Tube Removal Tool 361668 A CAUTION If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories Remove the rotor from the centrifuge by lifting it straight up and off the drive hub Return the rotor to the rotor vise Remove the plugs with the torque wrench To avoid stripping the plugs apply downward pressure to the hex plug adapter while loosening the plugs Use the appropriate removal tool see the SUPPLY LIST to remove the spacers and tubes VTi 50 Rotor TUBES The VTi 50 rotor uses only OptiSeal and Quick Seal tubes use only the tubes and accessories listed in Table 1 Refer to Rotors and Tubes for information on the chemical resistances of tube and accessory materials OptiSeal and Quick Seal tubes are disposable and should be discarded after a single use Table 1 Available Tubes for the VTi 50 Rotor Use only the items listed here Tube Required Accessory Max Speed Dimensions Part Part RCF and Volume Description Number Description Number Tube Rack k Factor 25x 89mm Quick Seal 344326 aluminum spacer 342417 348124 50 000 rom 39 mL Ultra Clear pkg 50 242 000 x g 36 25x 89mm Quick Seal 342414 aluminum spacer 342417 348124 50 000 rom 39 mL polyallomer pkg 50 242 000 x g 36 25 x 89 mm OptiSeal 362183 gold anodized 362204 360542 50 000 rom 36 2
14. Tube Topper kit 60 Hz 358312 Quick Seal Cordless Tube Topper kit 50 Hz Europe 358313 Quick Seal Cordless Tube Topper kit 50 Hz Great Britain 358314 Quick Seal Cordless Tube Topper kit 50 Hz Australia 358315 Quick Seal Cordless Tube Topper kit 50 Hz Canada 367803 Tube Topper rack 25 mm dia tubes 0 00000 348124 Torque wrench assembly 3 8 in drive 369791 Hex plussadapter ss enia ea tee eee ae eg 976959 Tube removal tool 0 0 eee eee een eee eee 361668 Floating spacer removal tool 00 0 0 eee eee eee 338765 Spinkote lubricant 1 0Z 0 0 0 cece eee eee 306812 Silicone vacuum grease 2 OZ o n eee cece eee eee 335148 Rotor Cleaning Kit 0 2 ee eee eee 339558 Beckman Solution 555 1 qt 0 ce eee eee eee 339555 Rotor cleaning brush 2 0 0 0 0 eee eee ee eee 339379 ULTRACENTRIFUGE ROTOR WARRANTY All Beckman Coulter ultracentrifuge Fixed Angle Vertical Tube Near Vertical Tube Swinging Bucket and Airfuge rotors are warranted against defects in materials or workmanship for the time periods indicated below subject to the Warranty Conditions stated below Preparative Ultracentrifuge Rotors 5 years No Proration Analytical Ultracentrifuge Rotors 5 years No Proration ML and TL Series Ultracentrifuge ROOTS eaen 0 esata e aon Cais Ree 5 years No Proration Airfuge
15. ach this notification will result in return or disposal of the items without review of the reported problem Use the address label printed on the RGA form when mailing the rotor and or accessories to Beckman Coulter Inc 1050 Page Mill Road Palo Alto CA 94304 Attention Returned Goods Customers located outside the United States should contact their local Beckman Coulter office 21 VTi 50 Rotor 22 SUPPLY LIST III NOTE To obtain copies of referenced publications contact Beckman Coulter Inc Technical Publi cations Department 1050 Page Mill Road Palo Alto CA 94304 U S A telephone 650 859 1753 fax 650 859 1375 Call Beckman Coulter Sales 1 800 742 2345 in the United States worldwide offices are listed on the back cover of this manual or see the Beckman Coulter Ultracentrifuge Rotors Tubes amp Accessories catalog BR 8101 for detailed information on ordering parts and supplies For your convenience a partial list is given below REPLACEMENT ROTOR PARTS OTHER VTi 50 rotor assembly 2 02 0 0 ee eee ee eee 362758 Rotor plug 2 6 A3ec don bashed he eid ah 355587 Rotor plus saskets isores serie AT E E E E N eas eas 340825 Overspeed disk 50 000 rpm 0 2 0 0 02 eee ee 330336 Rotor vise assembly 00 0 0 eee eee eine 332688 Tubes and accessories 0 cece eee eee see Table 1 OptiSeal tube rack assembly 00 00 0000 0 eee eee 360542 Quick Seal Cordless
16. ad 81410 Soganlik Kartal Istanbul Turkey Telephone 90 216 309 1900 Fax 90 216 309 0090 UNITED KINGDOM Beckman Coulter United Kingdom Ltd Oakley Court Kingsmead Business Park London Road High Wycombe Bucks HP11 1JU England U K Telephone 01494 441181 Fax 01494 447558 e mail beckman_uk beckman com Authorized dealers in other countries 4300 N Harbor Boulevard Box 3100 Fullerton California 92834 3100 Sales 1 800 742 2345 Service 1 800 551 1150 Internet www beckmancoulter com Telex 678413 Fax 1 800 643 4366 Cry ce Printed on recycled paper 2001 Beckman Coulter Inc amp LS EN ISO 9001
17. cept the drive unit which will be replaced at the then current price less a credit determined by the total number of revolutions or years completed provided that such a unit was manufactured or rebuilt by Beckman Coulter and ii if the centrifuge is cur rently covered by a Beckman Coulter warranty or Full Service Agreement all labor necessary for repair of the centrifuge 3 If a Beckman Coulter rotor covered by this warranty is dam aged due to a malfunction of a Beckman Coulter ultracentrifuge covered by an Ultracentrifuge System Service Agreement Beckman Coulter will repair or replace the rotor free of charge 4 If a Beckman Coulter rotor covered by this warranty is damaged due to a failure of a Beckman Coulter tube bottle tube cap spacer or adapter covered under the Conditions of this Warranty Beckman Coulter will repair or replace the rotor and repair the instrument as per the conditions in policy point 2 above and the replacement policy 5 Damage to a Beckman Coulter rotor or instrument due to the failure or malfunction of a non Beckman Coulter tube bottle tube cap spacer or adapter is not covered under this warranty although Beckman Coulter will assist in seeking compensation under the manufacturer s warranty Disclaimer IT IS EXPRESSLY AGREED THAT THE ABOVE WARRANTY SHALL BE IN LIEU OF ALL WARRANTIES OF FITNESS AND OF THE WARRANTY OF MERCHANTABILITY AND BECKMAN COULTER INC SHALL HAVE NO LIABILITY FOR SPE
18. d this would change the position of the sample bands Figure 2 together with the description and examples below show how to reduce run speeds when using CsCl gradients SELECTING CsCl GRADIENTS I gt NOTE The curves in Figure 2 are for solutions of CsCl salt dissolved in distilled water only If other salts are present in significant concentrations the overall CsCl concentration may need to be reduced Solid CsCl has a density of 4 g mL and if precipitation during centrifugation may cause rotor failure Precipitation will also alter density distribution and therefore sample separation In general lower speeds provide better resolution but longer run times will be required to achieve particle separation and gradient equilibrium Curves are provided up to the maximum rated speed of the rotor Rotor speed is used to control the slope of a CsCl density gradient and must be limited so that CsCl precipitation is avoided Figure 2a gives the CsCl concentration limiting curves for full tubes The refer ence curves in Figure 2b show equilibrium gradients that result from centrifugation using the maximum densities allowed by Figure 2a Each curve in Figure 2b is within the density limits allowed for the VTi 50 rotor each curve was generated for a single run speed using the maximum allowable homogeneous CsCl densities that avoid precipitation at that speed Figure 2c gives the gradients that result from centrifugation using lower than maxim
19. ients Up to 312 mL of gradient and sample can be centrifuged per run The rotor is made of titanium and is finished with black polyurethane paint A tube spacer and hex cavity rotor plug hold each tube in the rotor and a plug gasket forms a closure around each plug Rotor plugs are black anodized aluminum and spacers are clear anodized aluminum for Quick Seal tubes and gold anodized aluminum for OptiSeal tubes Because of the weight of the rotor drive pins are not required in the rotor drive hub cavity For overspeed protection a photoelectric detector in the ultracentri fuge monitors the overspeed disk on the rotor bottom and shuts down the run if speeds exceeding 50 000 rpm are detected See the Warranty at the back of this manual for warranty information PREPARATION AND USE Specific information about the VTi 50 rotor is given here Information common to this and other rotors is contained in Rotors and Tubes for Preparative Ultracentrifuges publication LR IM which should be used together with this manual for complete rotor and accessory operation Publication LR IM is included in the literature package with this rotor manual PRERUN SAFETY CHECKS AN 8 T A mse a W 830336 4 gt Can ROTOR PREPARATION VTi 50 Rotor I gt NOTE Although rotor components and accessories made by other manufacturers may fit in the VTi 50 rotor their safety in this rotor cannot be ascertained by Beckman Coulte
20. lled to the same level with liquid of the same density Figure 1 Arranging Tubes in the Rotor Two four six or eight tubes can be centrifuged per run if they are arranged in the rotor as shown 4 Complete loading by placing the correct spacers and floating spacers if applicable over the tubes It is important that each cavity being used is completely filled 5 Insert a rotor plug 355587 gasket end down over each spacer and screw it in I gt NOTE Do not use rotor plugs in empty cavities 6 Using the hex plug adapter 355588 and torque wrench 369791 tighten each rotor plug to 17 5 Nem 150 in Ib To avoid strip ping the plugs apply downward pressure to the hex plug adapter while tightening the plugs Do not overtighten plugs the top surface of each rotor plug should be flush with the surrounding rotor surface OPERATION VTi 50 Rotor Carefully place the rotor on the drive hub Refer to the instrument instruction manual for ultracentrifuge operation For additional operating information see the following RUN TIMES page 10 for using k factors to adjust run durations RUN SPEEDS page 11 for information about speed limitations SLOW ACCELERATION DECELERATION page 11 for informa tion about using slow acceleration and deceleration for gradient stability SELECTING CsCl GRADIENTS page 13 for methods to avoid CsCl precipitation during centrifugation REMOVAL AND SAMPLE RECOVERY
21. m allowable rotor speed is deter mined from the point where this curve intersects the homogeneous CsCl density 40 000 rpm 2 In Figure 2b sketch in a horizontal line corresponding to each particle s buoyant density 3 Mark the point in the figure where each particle density intersects the curve corresponding to the selected run speed and temperature 4 Particles will band at these locations across the tube diameter lower axis of Figure 2b at equilibrium during centrifugation After centrifugation the bands will reorient top axis If the required gradient curve is not presented in Figure 2b interpo late between the nearest curves and draw it in Using the horizontal axis it can be estimated that these particles will be about 5 mm apart at equilibrium during centrifugation In the 34 mL tube they will be separated by 9 4 mL top axis Example B Knowing particle buoyant densities 1 60 and 1 61 g mL how do you achieve the best separation 1 In Figure 2c sketch in a horizontal line corresponding to each particle s buoyant density 2 Select the curve at the temperature 20 C that gives the best particle separation Particles will band at points across the tube diameter where the sketched lines intersect this curve lower axis at equilibrium during centrifugation After centrifugation the bands will reorient top axis 17 VTi 50 Rotor Note the run speed along the selected curve Select the maximum h
22. nd dry thoroughly then lubricate lightly but evenly with Spinkote to coat all threads If the rotor or other components are contaminated with toxic or pathogenic materials follow appropriate decontamination procedures as outlined by your laboratory safety officer Check Appendix A in Rotors and Tubes to be sure the decontamination method will not damage any part of the rotor 19 VTi 50 Rotor 20 STERILIZATION AND DISINFECTION e The rotor and all rotor components can be autoclaved at 121 C for up to an hour Remove the plugs from the rotor and place the rotor oe plugs and spacers in the autoclave upside down Ethanol 70 or hydrogen peroxide 6 may be used on all rotor components including those made of plastic Bleach sodium hypochlorite may be used but may cause discoloration of anodized surfaces Use the minimum immersion time for each solution per laboratory standards Guam ut While Beckman Coulter has tested these methods and found that they do not damage the rotor or components no guarantee of sterility or disinfection is expressed or implied When sterilization or disinfec tion is a concern consult your laboratory safety officer regarding proper methods to use OptiSeal and Quick Seal tubes are disposable and should be discarded after a single use STORAGE When it is not in use store the rotor in a dry environment not in the instrument with plugs removed to allow air circulation so moistu
23. ns largest tube 25 x 89 mm Nominal tube capacity 0 0 eee eee eee 39 mL Nominal rotor capacity 0 2 2 ee eee 312 mL Approximate acceleration time to maximum speed fully loaded 0 cece 20 min Approximate deceleration time from maximum speed fully loaded 0 0 2 0 0 2c eee eee ee 14 min Weight of fully loaded rotor 0 0 e eee eee 12 kg 27 lb Rotor material 0 2 2 eee eee titanium Relative Centrifugal Field RCF is the ratio of the centrifugal acceleration at a specified radius and speed r to the standard acceleration of gravity g according to the following formula RCF 1 amp 2 amp where r is the radius in millimeters is the angular velocity in radians per second 2 7 RPM 60 and g is the standard acceleration of gravity 9807 mm s2 After substitution 2 RCF 1 12r Fa 1000 VTi 50 Rotor DESCRIPTION Plug Plug lt 355587 Rotor Gasket Bod aen E a ad N Overspeed Disk 330336 This rotor has been manufactured in an NSAI registered ISO 9001 or 9002 facility for use with the specified Beckman Coulter ultracentrifuges The VTi 50 rated for 50 000 rpm is designed to centrifuge up to eight tubes in an upright position Used in Beckman Coulter class H R and S preparative ultracentrifuges the rotor develops centrifugal forces that can efficiently band DNA or isolate proteins on density grad
24. omogeneous CsCl density that corresponds to the temperature and run speed established above These param eters will provide the particle banding pattern selected in Step 2 In this case the 1 6 g mL 30 000 rpm curves in Figure 2c give the best separation These curves intersect the particle buoyant densities in such a way that particles band about 2 mm apart at equilibrium during centrifugation lower axis Bands are located in the middle of the tube CARE AND MAINTENANCE MAINTENANCE Centering gt NOTE Tool Do not use sharp tools on the rotor that could 991355 A cause scratches in the rotor surface Corrosion Plug p Plug Gasket 355587 340825 E Check for Corrosion N Overspeed Disk 330336 18 begins in scratches and may open fissures in the rotor with continued use Regularly inspect the overspeed disk If it is scratched damaged or missing replace it Replacement instructions are in Rotors and Tubes Regularly inspect the rotor plugs 355587 for wear worn threads will have a shiny appearance Replace worn plugs Regularly lubricate the metal threads in the rotor plugs with a thin even coat of Spinkote lubricant Failure to keep these threads lubri cated can result in damaged threads Replace rotor plugs as a set if they show signs of wear The rotor plug gaskets 340825 require no maintenance except cleaning Replace damaged gaskets To replace the plug gasket use the sharpened end
25. or heat With the tube spacer and rotor plug in place the combination of g force and hydrostatic pressure during centrifugation ensures a tight reliable seal that protects your samples Fill each tube to the base of the stem leaving no fluid in the stem Overfilling the tube can cause spillage when the plug is inserted or compromise seal integrity however too much air can cause the tube to deform disrupting gradients and sample bands Refer to Using OptiSeal Tubes publication IN 189 included in each package of OptiSeal tubes for detailed information on the use and care of OptiSeal tubes Quick Seal Tubes Quick Seal tubes must be sealed prior to centrifugation These tubes are heat sealed and do not need caps however spacers are required on top of the tubes when they are loaded into the rotor e Fill Quick Seal tubes leaving a small bubble of air at the base of the neck Do not leave a large air space too much air can cause excessive tube deformation and make the tube difficult to remove e Some of the Quick Seal tubes listed in Table 1 are part of the g Max system which uses a combination of small bell top Quick Seal tubes and floating spacers also called g Max spacers This means that you can run the shorter tubes in this rotor without reduction in g force For detailed information on the g Max system see publication DS 709 Refer to Rotors and Tubes for detailed information on the use of Quick Seal tubes VTi 50
26. r Use of other manufacturers components or accessories in the VTi 50 rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in this publication should be used in this rotor Read the Safety Notice page at the front of this manual before using the rotor 1 Inspect the rotor plugs and gaskets for damage the high forces generated in this rotor can cause damaged components to fail 2 Make sure that the rotor is equipped with the correct overspeed disk If the disk is missing or damaged replace it according to the instructions in Rotors and Tubes 3 Verify that only the tubes and accessories listed in Table 1 are being used Check the chemical compatibilities of all materials used refer to Appendix A in Rotors and Tubes For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration 1 Be sure that the plug threads are clean and lightly but evenly lubri cated with Spinkote lubricant 306812 to ensure a proper seal by minimizing thread friction 2 Set the rotor in the rotor vise 332688 which should be bolted or clamped to a rigid surface VTi 50 Rotor 3 Load the filled and plugged or sealed tubes symmetrically into the rotor see page 8 for tube information If fewer than eight tubes are being run they must be arranged symmetrically in the rotor see Figure 1 Opposing tubes must be fi
27. re will not collect in the tube cavities Flammability hazard Do not use in or near operating ultracentrifuges VTi 50 Rotor RETURNING A ROTOR Before returning a rotor or accessory for any reason prior permission RGA a Returned Goods Authorization form must be obtained from 2 Beckman Coulter Inc This RGA form may be obtained from your _ local Beckman Coulter sales office and should contain the following ee ja information e serial number e history of use approximate frequency of use e reason for the return e original purchase order number billing number and shipping number if possible e name and phone number of the person to be notified upon receipt of the rotor or accessory at the factory e name and phone number of the person to be notified about repair costs etc To protect our personnel it is the customer s responsibility to ensure that all parts are free from pathogens and or radioactivity Steriliza tion and decontamination must be done before returning the parts Smaller items such as tubes bottles etc should be enclosed in a sealed plastic bag All parts must be accompanied by a note plainly visible on the out side of the box or bag stating that they are safe to handle and that they are not contaminated with pathogens or radioactivity Failure to att
28. sed select slow acceleration as follows ACCEL e in Optima XL series ultracentrifuges select a slow acceleration PROFILES profile e in Optima L series ultracentrifuges select SLOW ACCEL For the stability of all gradients during deceleration do the following DECEL e in Optima XL series ultracentrifuges select a slow deceleration PROFILES profile e in Optima L series ultracentrifuges select SLOW DECEL e in L7 series ultracentrifuges set the brake switch in the 800 rpm position RUN SPEEDS The centrifugal force at a given radius in a rotor is a function of SPEED RPMRCF speed Comparisons of forces between different rotors are made by comparing the rotors relative centrifugal fields RCF When rota Sa SEE RAT tional speed is adjusted so that identical samples are subjected to the same RCF in two different rotors the samples are subjected to the same force The RCF at a number of rotor speeds is provided in Table 2 Speeds must be reduced under the following circumstances 1 If nonprecipitating solutions more dense than 1 7 g mL are centri fuged the maximum allowable run speed must be reduced according to the following equation reduced maximum speed 50 000 rpm ra 6 where p is the density of the tube contents This speed reduction will protect the rotor from excessive stresses due to the added tube 11 VTi 50 Rotor Table 2 Relative Centrifugal Fields for the VTi 50 Rotor 5 1 mL Tubes En
29. stemic fungi further emphasize the need for aerosol protection Handle other infectious samples according to good laboratory procedures and methods to prevent spread of disease Because spills may generate aerosols observe proper safety precautions for aerosol contain ment Do not run toxic pathogenic or radioactive materials in this rotor without taking appropriate safety precautions Biosafe containment should be used when Risk Group II materials as identified in the World Health Organization Laboratory Biosafety Manual are handled materials of a higher group require more than one level of protection The rotor and accessories are not designed for use with materials capable of developing flammable or explosive vapors Do not centrifuge such materials in nor handle or store them near the ultracentrifuge Although rotor components and accessories made by other manufacturers may fit in the VTi 50 rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the VTi 50 rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in this publication should be used in this rotor Do not run an empty rotor Place filled tubes in at least two opposing cavities Make sure that filled containers are loaded symmetrically into the rotor and that opposing tubes are filled to the same level with liquid of the
30. tries in this table are calculated from the formula RCF 1 12r RPM 1000 and then rounded to three significant digits Relative Centrifugal Field x g Rotor Speed Aa At fav A k rom 86 6 mm 73 7 mm 60 8 mm Factor 50 000 242 000 206 000 170 000 36 45 000 196 000 167 000 138 000 44 40 000 155 000 132 000 109 000 56 35 000 119 000 101 000 83 400 T3 30 000 87 300 74 300 61 300 100 25 000 60 600 51 600 42 600 143 20 000 38 800 33 000 27 200 224 15 000 21 800 18 600 15 300 398 10 000 9 700 8 250 6 800 896 5 000 2 430 2 060 1700 3 584 Calculated for all Beckman Coulter preparative rotors as a measure of the rotor s relative efficiency in pelleting sample in water at 20 C Relative Centrifugal Fields for the VTi 50 Rotor 300 000 250 000 200 000 150 000 100 000 20 000 Speed rpm 12 VTi 50 Rotor load Note however that the use of this formula may still produce maximum speed figures that are higher than the limitations imposed by the use of certain tubes or adapters In such cases use the lower of the two figures 2 Further speed limits must be imposed when CsCl or other self forming gradient salts are centrifuged as equation 6 does not predict concentration limits speeds that are required to avoid precipitation of salt crystals Precipitation during centrifugation would alter the density distribution of CsCl an
31. um allowable CsCl concentrations These reduced density curves can be used to make particles band more towards the middle of a tube where volume 13 VTi 50 Rotor 1 9 1 8 1 7 fay e E z Q 5 O op 5 45 ic a 5 e o D O 1 4 1 3 1 2 1 4 VTi 50 ROTOR 20 C 4 C 10 20 30 40 50 Rotor Speed K rpm Figure 2a Precipitation Curves for the VTi 50 Rotor Using combinations of rotor speeds and homogeneous CsCl solution densities that intersect on or below these curves ensures that CsCl will not precipitate during centrifugation If gradient and sample solutions do not completely fill the tube add mineral oil to fill Do not use an oil overlay in Ultra Clear tubes The dashed line is a representation of equation 6 and is shown here to illustrate the inability of that equation to predict CsCl precipitation 14 VTi 50 Rotor AT EQUILIBRIUM During or After Centrifugation
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