Instruction Manual
Contents
1. 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 150 Rotor Speed x1000 RPM 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 PN 393554AA 15 MLA 150 Rotor Selecting CsCl Gradients Figure 4 CsCl Gradients at Equilibrium for the MLA 150 Rotor with Quick Seal Tube 344625 1 7 7 7 1 8 i i l i t 1 7 Pt 50 000 i L 70 000 l 1 6 Pe ee eet e a A i E15 rt 90 000 2 oo 000 S z al Cr 1 3 1 2 MLA 150 ROTOR 2c i 4C l 4 16 18 20 22 24 26 28 30 32 34 36 38 40 Distance from Axis of Rotation mm Centrifugation of homogeneous CsCl solutions at the maximum allowable speeds from Figure 3 results in gradients presented in Figure 4 16 PN 393554AA MLA 150 Rotor Care and Maintenance Care and Maintenance2. Maintenance 4 Lid Assembly Threads we e N Lid o ring 860218 SS Check for Corrosion NOTE 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 e Regularly lubricate the metal threads in the rotor with a thin even coat of Spinkote lubricant Failure to keep these threads lubricated can result in damaged threads e Regularly apply silicone vacuum grease to the lid O ring Replace the O ring about twice a year or whenever worn or damaged This lid O ring 860218 is specifically designed for use with the MLA 150 rotor and is not interchangeable with other O rings Do not substitute any other O ring for use in this rotor e Check lid center threads for evidence of distortion overtightening or damage Refer to Appendix A in Rotors and Tubes for Tabletop Preparative Ultracentrifuges for the chemical resistances of rotor and accessory materials Your Beckman Coulter representative is your point of contact for the Field Rotor Inspection Program and the rotor repair center PN 393554AA MLA 150 Rotor Care and Maintenance Cleaning Rotor Cleaning Kit 339558 a 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 clea
3. Tube Topper rack Tube removal tool Curved hemostat 6 in Straight hemostat 6 in Fraction Recovery System Fraction Recovery System Adapter Kit for TL series tubes Spinkote lubricant 2 oz Silicone vacuum grease 1 oz Rotor Cleaning Kit Rotor cleaning brush Beckman Solution 555 1 qt 393489 860218 343778 347287 344624 344625 393570 358312 367803 358313 358314 358315 349387 361668 927208 961519 342025 347828 306812 335148 339558 347404 339555 21 PN 393554AA 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 Rotors 5 years No Proration Airfuge 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 8 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 This warranty extends only to the original Buyer and may not be assigned or extended to a third pers
4. reduce the maximum allowable run speed according to the following equation reduced maximum speed 150 000 RPM 7 g ml a 6 where p is the density g mL of the tube contents This speed reduction will protect the rotor from excessive stresses due to the added tube load Note however that the use of this formula may still produce maximum speed values that are higher than the limitations imposed by the use of certain tubes or adapters In such cases use the lower of the two values e You must impose further speed limits 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 and this would change the position of the sample bands The illustrations and examples in Selecting CsCl Gradients show how to reduce run speeds when using CsCl gradients MLA 150 Rotor Preparation and Use Table 2 Relative Centrifugal Fields for Cell Holes of the MLA 150 Rotor ee Relative Centrifugal Field x g k Factor Atr max 39 9 mm At r av 27 8 mm Atr min 15 8 mm 150 000 1 003 000 700 000 397 000 10 4 145 000 937 000 654 000 371 000 11 140 000 874 000 610 000 346 000 12 135 000 812 000 567 000 322 000 13 130 000 753 000 526 000 298 000 14 125 000 697 000 486 000 276 000 15 120 000 642 000 448 000 254 000 16 115 000 590 000
5. 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 Europark Fichtenhain B 13 47807 Krefeld Germany Telephone 49 21 513335 Fax 49 21 51333633 e mail bioresearch de beckman com GS zraou COULTER Beckman Coulter Inc Sales and Service 1 800 742 2345 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 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 JAPAN Beckman Coulter K K TOC Ariake West Tower 2 5 7 Ariake Koto ku Tokyo 135 0063 Japan Telephone 03 5404 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 Mexico D F Mexico Telephone 52 55 9183 2800 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 6339 3633 F
6. Box 3100 Fullerton California 92834 3100 e Fax 1 800 643 4366 G Bd Printed on recycled paper
7. Factor calculations use rmax values that are reduced by the tube wall thickness b Ultem is a registered trademark of GE Plastics MLA 150 Rotor Preparation and Use Quick Seal Tubes Spacer Ca A aa Bell top Tube 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 e Refer to Rotors and Tubes for Tabletop Preparative Ultracentrifuges for detailed information on the use and care of Quick Seal tubes Thickwall Tubes J Thickwall polyallomer and polycarbonate tubes can be run partially filled maximum of 1 ml without caps All opposing tubes installed in a rotor must be filled with a like substance to maintain balance during centrifugation To avoid spillage do not overfill capless tubes Be sure to note reductions in centrifugation speed shown in Table 1 Run Times The k factor of the rotor is a measure of the rotor s pelleting efficiency Beckman Coulter has calculated the k factors for all of its preparative rotors at maximum rated speed using tubes filled to their maximum recommended levels The k factor is calculated from the formula 1n Tmax Tmin 10 3 k 2 where o is the angular ve
8. 412 000 234 000 18 110 000 539 000 377 000 214 000 19 105 000 491 000 343 000 195 000 21 100 000 446 000 311 000 177 000 23 95 000 402 000 281 000 159 000 26 90 000 361 000 252 000 143 000 29 85 000 322 000 225 000 128 000 32 80 000 285 000 199 000 113 000 37 75 000 251 000 175 000 99 300 42 70 000 218 000 152 000 86 500 48 65 000 188 000 131 000 74 600 56 60 000 160 000 112 000 63 600 65 55 000 135 000 94 100 53 400 78 50 000 111 000 77 800 44 200 94 45 000 90 300 63 000 35 800 116 40 000 71 300 49 800 28 300 147 35 000 54 600 38 100 21 600 191 30 000 40 100 28 000 15 900 261 25 000 27 900 19 400 11 000 375 20 000 17 800 12 400 7 060 586 15 000 10 000 7 000 3 970 1 042 10 000 4 460 3 110 1770 2 345 5 000 1110 778 442 9 380 digits water at 20 C Entries in this table are calculated from the formula RCF 1 12r RPM 1 000 2 The r values are rounded to three significant Calculated for all Beckman Coulter preparative rotors as a measure of the rotor s relative efficiency in pelleting sample in PN 393554AA MLA 150 Rotor Selecting CsCl Gradients Figure 2 Relative Centrifugal Fields RCF for the MLA 150 Rotor MLA 150 Relative Centrifugal Fields RCF Cell Hole 1 200 000 1 000 000 800 000 D amp 600 000 O ao 400 000 200 000 0 0 20 000 40 000 60 000 80 000 100 000 120 000 140 000 160 000 Speed rpm Selecting CsCl Gradients PN 393554AA Precipitation during centrifug
9. PN 393554AA August 2007 Beckman Coulter Inc 4300 N Harbor Blvd Fullerton CA 92835 Instruction Manual MLA 150 Fixed Angle Rotor Used in the Beckman Coulter Optima MAX XP Ultracentrifuge lt BECKMAN COULTER o MLA 150 Rotor PN 393554AA August 2007 Copyright 2007 Beckman Coulter Inc All rights reserved No part of this document may be reproduced or transmitted in any form or by any means electronic mechanical photocopying recording or otherwise without prior written permission from Beckman Coulter Inc Find us on the World Wide Web at www beckmancoulter com Safety Notice Safety Notice PN 393554AA 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 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 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 Co
10. Ultracentrifuge System Service Agreement Beckman Coulter will repair or replace the rotor free of charge Ultracentrifuge Rotor Warranty 4 Ifa 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 SPECIAL OR CONSEQUENTIAL DAMAGES OF ANY KIND WHATSOEVER ARISING OUT OF THE MANUFACTURE 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 centrif
11. ample particles Typical Examples for Determining CsCl Run Parameters Example A A separation that is done frequently is the banding of plasmid DNA in cesium chloride with ethidium bromide The starting density of the CsCl solution is 1 55 g mL In this separation the covalently closed circular plasmid bands at a density of 1 57 g mL while the nicked and linear species band at 1 53 g mL At 20 C where will particles band At Speed At Rest in Rotor Upright Floating Components Bands Pelleted Material 1 In Figure 3 find the curve that corresponds to the desired run temperature 20 C and fill volume in this case full The maximum allowable rotor speed 97 300 RPM is determined from the point where this curve intersects a line drawn at the homogeneous CsCI density 1 55 g mL 2 In Figure 4 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 90 000 RPM and temperature 20 C 4 Particles will band at these locations across the tube diameter at equilibrium during centrifugation In this example particles will band about 28 0 and 26 1 mm from the axis of rotation about 1 9mm of centerband to centerband separation at the rotor s 30 degree tube angle When you hold the tube upright there will be about 2 2 mm of centerband to centerband separation You can calculate this in
12. ation would alter density distribution and this would change the position of the sample bands Curves in Figure 3 and Figure 4 are provided up to the maximum rated speed of the rotor The curves in Figure 3 and Figure 4 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 Rotor speed is used to control the slope of a CsCl density gradient and must be limited to avoid CsCl precipitation Speed and density combinations that intersect on or below the curves in Figure 3 MLA 150 Rotor Selecting CsCl Gradients ensure that CsCl will not precipitate during centrifugation in the MLA 150 rotor Curves are provided at two temperatures 20 C black curves and 4 C gray curves The reference curves in Figure 4 show gradient distribution at equilibrium Each curve in Figure 4 is within the density limits allowed for the MLA 150 rotor each curve was generated for a single run speed using the maximum allowable homogeneous CsCl densities one for each fill level that avoid precipitation at that speed The gradients in Figure 4 can be generated from step or linear gradients or from homogeneous solutions However the total amount of CsCl in solution must be equivalent to a homogeneous solution corresponding to the concentrations specified in Figure 3 Figure 4 can also be used to approximate the banding positions of s
13. ax 65 6336 6303 SOUTH AFRICA SUB SAHARAN 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 e mail beckman intekom co za Internet www beckmancoulter com Telex 678413 2007 Beckman Coulter Inc SPAIN Beckman Coulter Espa a S A C Caleruega 81 28033 Madrid Spain Telephone 91 3836080 Fax 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 Cad 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 Buckinghamshire 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
14. e reported problem Use the address label printed on the RGA form when mailing the rotor and or accessories to Beckman Coulter 1050 Page Mill Road Palo Alto CA 94304 Attention Returned Goods Customers located outside the United States should contact their local Beckman Coulter office PN 393554AA Supply List MLA 150 Rotor Supply List See the Beckman Coulter Ultracentrifuge Rotors Tubes amp Accessories catalog BR 8101 or contact Beckman Coulter Sales 1 800 742 2345 in the United States worldwide offices are listed on the back cover of this manual for detailed information on ordering parts and supplies For your convenience a partial list is given below Publications referenced in this manual can be obtained by calling 1 800 742 2345 in the United States or by contacting your local Beckman Coulter office Replacement Rotor Parts Other PN 393554AA MLA 150 rotor assembly Lid O ring Tubes and accessories Open Top Tube 1 0 ml Polycarbonate Package of 100 Open Top Tube 1 0 ml Polyallomer Package of 100 Quick Seal Tube 1 5 ml Polyallomer Package of 50 Quick Seal Tube 2 0 ml Polyallomer Package of 50 Spacer for Quick Seal Tube Package of 8 Quick Seal Cordless Tube Topper kit 60 Hz Quick Seal Cordless Tube Topper kit 60 Hz Canada Quick Seal Cordless Tube Topper kit 50 Hz Europe Quick Seal Cordless Tube Topper kit 50 Hz Great Britain Quick Seal Cordless Tube Topper kit 50 Hz Australia
15. er devices not of Beckman Coulter manufacture 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 bearings seals and lubrication are excluded from this warranty and should be frequently inspected and replaced if they become worn or damaged 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 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 except 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 currently covered by a Beckman Coulter warranty or Full Service Agreement all labor necessary for repair of the centrifuge If a Beckman Coulter rotor covered by this warranty is damaged due to a malfunction of a Beckman Coulter ultracentrifuge covered by an
16. lable tubes See Table 1 Nominal tube dimensions largest volume tube 344625 11 x 32 mm PN 393554AA 5 MLA 150 Rotor Description Nominal tube capacity largest volume tube 344625 2 mL Nominal rotor capacity 16 mL Approximate acceleration time to maximum speed fully loaded 3 min Approximate deceleration time from maximum speed fully loaded 3 min Weight of fully loaded rotor 0 40 kg 0 89 1b Rotor material titanium Lid material aluminum Relative Centrifugal Field RCF is the ratio of the centrifugal acceleration at a specified radius and speed re to the standard acceleration of gravity g according to the following formula RCE 2 g where r is the radius in millimeters is the angular velocity in radians per second 2 n RPM 60 and g is the standard acceleration of gravity 9807 mm s After substitution RPMN RCF 1 12r 1000 Description Lid Assembly Z o Lid O ring 860218 This rotor has been manufactured in a registered ISO 9001 or 13485 facility for use with the appropriately classified Beckman Coulter ultracentrifuge The MLA 150 rotor rated for 150 000 RPM has a tube angle of 30 degrees from the axis of rotatio
17. locity of the rotor in radians per second 0 105 x RPM max is the maximum radius and r yi is the minimum radius After substitution 11 2 533 x 10 1n Tmax Tmin 2 RPM 2 PN 393554AA MLA 150 Rotor Preparation and Use Use the k factor in the following equation to estimate the run time t in hours required to pellet particles of known sedimentation coefficient s in Svedberg units S 5 3 Run times can be estimated for centrifugation at less than maximum speed by adjusting the k factor as follows back 150000 2 adj actual run speed 4 Run times can also be estimated from data established in prior experiments if the k factor of the previous rotor is known For any two rotors a and b ta _ ka th kp For more information on k factors see Use of k Factor for Estimating Run Times from Previously Established Run Conditions publication DS 719 Run Speeds PN 393554AA The centrifugal force at a given radius in a rotor is a function of speed You can compare forces between different rotors by comparing the rotors relative centrifugal fields RCF Identical samples in different rotors are subjected to the same force if the speed of the rotors is adjusted to produce the same RCF The RCF for the cell hole and various labware is provided in Table 2 You must reduce speeds under the following circumstances e Ifyoucentrifuge non precipitating solutions having a density greater than 1 7 g mL
18. n The rotor can centrifuge up to eight tubes and is used in the Beckman Coulter Optima MAX XP ultracentrifuge The rotor is made of titanium and is finished with black polyurethane paint The lid is made of aluminum and is anodized to resist corrosion A rotor retention mechanism on the ultracentrifuge drive hub secures the rotor during the run A lubricated O ring in the lid maintains atmospheric pressure inside the rotor during centrifugation The tube cavities are numbered to aid in sample identification 6 PN 393554AA MLA 150 Rotor Preparation and Use The ultracentrifuge identifies rotor speed during the run by means of a magnetic speed sensor in the instrument chamber and magnets on the bottom of the rotor This overspeed protection system ensures that the rotor does not exceed its maximum speed See the Warranty at the back of this manual for warranty information Preparation and Use Specific information about the MLA 150 rotor is given here Information common to this and other rotors is contained in Rotors and Tubes for Tabletop Preparative Ultracentrifuges publication TLR IM which should be used with this manual for complete rotor and accessory operation Publication TLR IM is included in the literature package shipped with the rotor Other referenced publications are available at www beckmancoulter com NOTE Although rotor components and accessories made by other manufacturers may fit in the MLA 150 rotor their
19. n the rotor frequently at least weekly to prevent buildup of residues Do not wash rotor components in a dishwasher Do not soak in detergent solution for long periods such as overnight 1 Remove the lid O ring before washing 2 Wash the rotor and lid in a mild detergent such as Beckman Solution 555 339555 that won t damage the rotor The Rotor Cleaning Kit 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 3 Rinse the cleaned rotor and components with distilled water 4 Air dry the rotor and lid upside down Do not use acetone to dry the rotor Ensure that magnet locations on the rotor bottom are thoroughly dry to prevent corrosion 5 Apply a thin even coat of silicone vacuum grease to the lid O ring before replacing it in the groove in the lid Clean metal threads as necessary at least every 6 months Use a brush and concentrated Solution 555 Rinse and dry thoroughly then lubricate lightly but evenly with Spinkote to coat all threads Lid Assembly Lid O ring x 860218 Periodically remove the lid O ring and wipe clean as necessary Clean the O ring groove with a cot
20. ollect in the tube cavities Flammability hazard Do not use in or near operating ultracentrifuges MLA 150 Rotor Returning a Rotor Returning a Rotor 20 Before returning a rotor or accessory for any reason prior permission a Returned Goods Authorization form must be obtained from Beckman Coulter Inc This RGA form may be obtained from your local Beckman Coulter sales office and should contain the following information 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 pathogen s and or radioactivity Sterilization 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 outside of the box or bag stating that they are safe to handle and that they are not contaminated with pathogens or radioactivity Failure to attach this notification will result in return or disposal of the items without review of th
21. on without written consent of Beckman Coulter 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 This warranty is void if the Beckman Coulter Centrifuge System 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 maintenance procedures centrifuge and rotor classification requirements 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 Rotor bucket 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 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 if the rotor is used with carriers buckets belts or oth
22. safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the MLA 150 rotor may void the rotor warranty and should be prohibited by your laboratory safety officer Only the components and accessories listed in this publications should be used in this rotor Prerun Safety Checks Read the Safety Notice page at the front of this manual before using the rotor 1 Inspect the O ring and lid for damage the high forces generated in this rotor can cause damaged components to fail 2 Make sure you use only tubes and accessories listed in Table 1 3 Check the chemical compatibilities of all materials used refer to Appendix A in Rotors and Tubes for Tabletop Preparative Ultracentrifuges Rotor Preparation For runs at other than room temperature refrigerate or warm the rotor beforehand for fast equilibration 1 Lightly but evenly lubricate metal threads with Spinkote lubricant 306812 2 Apply a thin film of silicone vacuum grease 335148 to the O ring in the rotor lid 3 Load the filled and capped tubes symmetrically into the rotor see Quick Seal Tubes If fewer than eight tubes are being run they must be arranged symmetrically in the rotor see Figure 1 Arranging Tubes in the Rotor Opposing tubes must be filled to the same level with liquid of the same density Two four six or eight tubes can be centrifuged per run if they are arranged in the rotor as
23. shown in Figure 1 PN 393554AA 7 MLA 150 Rotor Preparation and Use Figure 1 Arranging Tubes in the Rotor 4 Use the required spacers if necessary see Table 1 to complete the loading operation 5 After loading the rotor place the lid onto the rotor threads Advance the lid to the right clockwise by hand until the O ring begins to contact the rotor body Continue tightening the lid approximately an additional 1 8 of a revolution No tool is required The distance between tube cavities is 1 8 of a revolution You can use the eight scallops in the rotor body as a visual guide Operation 1 Use an absorbent towel to wipe off condensation from the rotor 2 Carefully place the rotor on the drive hub 3 For information on ultracentrifuge operation refer to the instrument instruction manual 4 For additional operating information see the following e Run Times for using k factors to adjust run durations e Run Speeds for information on speed limitations e Selecting CsCl Gradients for methods to avoid CsCl precipitation during centrifugation Removal and Sample Recovery If disassembly reveals evidence of leakage you should assume that some fluid escaped from the rotor Apply appropriate decontamination procedures to the centrifuge and accessories 1 Remove the rotor from the ultracentrifuge and place it on a flat surface 2 Remove the lid by unscrewing it to the left counterclockwise 3 Use a
24. terband distance dup from the formula PN 393554AA MLA 150 Rotor Selecting CsCl Gradients where dg is the interband distance when the tube is held at an angle 9 in the rotor Example B Knowing particle buoyant densities e g 1 59 and 1 54 g mL how do you achieve good separation 1 InFigure 4 sketch in a horizontal line corresponding to each particle s buoyant density 2 Select the 70 000 RPM curve at the desired temperature 4 C to give the best particle separation that is a shallow gradient with the material positioned near the middle of the tube 3 With arun speed of 70 000 RPM and a selected fill volume full determine the maximum homogeneous CsCI density from Figure 3 4 From Figure 3 the maximum homogeneous CsCl density 1 637 g mL is found by locating the point on the Full curve for 4 C where it crosses 70 000 RPM Using a homogenous CsCI density of 1 55 g mL will provide the particle banding pattern selected in Step 2 In this example particles will band at about 25 6 and 21 3 mm from the axis of rotation about 2 3 mm apart When you hold the tube upright there will be about 2 7 mm of center of band to center of band separation Figure 3 CsCl Precipitation Curves for the MLA 150 Rotor SS Pe a MLA 150 ROTOR 20 C Homogeneous CsCl Solution g ml 4 C
25. ton tipped swab Reapply a light film of silicone vacuum grease PN 393554AA MLA 150 Rotor Care and Maintenance Decontamination pia If the rotor or other components are contaminated with radioactive toxic or pathogenic materials follow appropriate decontamination procedures as outlined by appropriate laboratory safety guidelines and or other regulations Consult Appendix A in Rotors and Tubes for Tabletop Preparative Ultracentrifuges to select an agent that will not damage the rotor Sterilization and Disinfection Storage PN 393554AA e The rotor and all rotor components can be autoclaved at 121 C for up to an hour Remove the lid and O ring from the rotor and place the rotor and lid in the autoclave upside down e 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 While Beckman Coulter has tested these methods and found that they do not damage the rotor components no guarantee of sterility or disinfection is expressed or implied When sterilization or disinfection is a concern consult your laboratory safety officer regarding proper methods to use When it is not in use store the rotor in a dry environment not in the instrument with the lid removed to allow air circulation so moisture will not c
26. tory safety officer Only the components and accessories listed in this publication should be used in this rotor 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 same density Make sure that cavities in use have the proper spacers and or floating spacers inserted before installing the rotor lid If disassembly reveals evidence of leakage you should assume that some fluid escaped the rotor Apply appropriate decontamination procedures to the centrifuge and accessories Never exceed the maximum rated speed of the rotor and labware in use Refer to 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 MLA 150 Rotor Introduction max so Axis of rotation gt Specifications Maximum speed 150 000 RPM Density rating at maximum speed 1 7 g mL Relative Centrifugal Field at maximum speed At Tmax 39 9 mm 1 003 000 x g max At ray 27 8 mm 700 200 x g At f min 15 8 mm 397 400 x g k factor at maximum speed 10 4 Conditions requiring speed reductions See Run Speeds Number of tube cavities 8 Avai
27. tube removal tool to remove the spacers and tubes 8 PN 393554AA Tubes and Accessories MLA 150 Rotor Preparation and Use The MLA 150 rotor uses tubes and accessories listed in Table 1 Be sure to use only those items listed and to observe the maximum fill volumes and speed limits shown Refer to Appendix A in Rotors and Tubes for Tabletop Preparative Ultracentrifuges for information on the chemical resistances of tube and accessory materials Temperature Limits 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 If plastic containers are frozen before use make sure that they are thawed to at least 2 C prior to centrifugation Table 1 Available Tubes for the MLA 150 Rotor PN 393554AA Tube Required Accessory Max Speed anes x RCF Dimensions Description Part Number Description Part Number k Factor Volume 11x 32mm Quick Seal 344625 Floating 393570 150 000 RPM 2 0 ml polyallomer pkg 50 Spacer Ultem gt pkg 8 989 000 xg bell top 8 0 11x 25mm Quick Seal 344624 Floating 393570 150 000 RPM 1 5 ml polyallomer pkg 50 Spacer Ultem pkg 8 989 000 xg bell top 6 2 11x 34mm Thickwall 343778 none 150 000 RPM 1 0 ml polycarbonate pkg 100 964 000 xg 4 6 11x 34mm Thickwall 347287 none 95 000 RPM 1 0 ml polyallomer pkg 100 387 000 xg 11 4 a The RCF and k
28. uge 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 ACCESSORIES ARE NON RADIOACTIVE NON PATHOGENIC NON TOXIC AND OTHERWISE SAFE TO SHIP AND HANDLE IS REQUIRED PN 393554AA Beckman Coulter Worldwide Biomedical 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 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 NORTH 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
29. ulter ultracentrifuge that has been modified without Beckman Coulter s approval Handle body fluids with care because they can transmit disease No known test offers complete assurance that they are free of micro organisms Some of the most virulent Hepatitis B and C viruses HIV I V viruses atypical mycobacteria and certain systemic 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 containment 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 MLA 150 rotor their safety in this rotor cannot be ascertained by Beckman Coulter Use of other manufacturers components or accessories in the MLA 150 rotor may void the rotor warranty and should be prohibited by your labora
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