User Manual
Contents
1. ORDERING INFORMATION To order The SHREDDER SG3 or Shredder Tubes and other accessories please contact your local Pres sure BioSciences Sales Representative Or please visit http www pressurebiosciences com Pressure BioSciences Inc 14 Norfolk Ave South Easton MA 02375 TEL 508 230 1828 FAX 508 230 1829 www pressurebiosciences com Pressure BioSciences Inc Discovery Starts With Sample Preparation The SHREDDER SG3 Gentle Rapid and Safe Disruption of Tissues and Organisms for the Extraction of DNA RNA Protein Mitochondria Lipid and Small Molecules User Manual a g A SAK For Research Use Only Not for Use in Diagnostic Procedures Barocycler PULSE Tubes The PCT Shredder The SHREDDER SG3 PCT Shredder PULSE Tubes and Shredder Tubes are trademarks of Pressure BioSciences Inc Table of Contents Safety Information Product Use Limitations Product Warranty and Satisfaction Guarantee Technical Assistance Introduction Kit Components Varieties of Shredder Tubes Applications Requiring PCT Preparing The SHREDDER SG3 How to Use The SHREDDER SG3 Protocols for The SHREDDER SG3 Application Notes and PrEPs Ordering Information 14 15 16 New Procedures are added periodically to the PBI Website APPLICATIONS NOTES AND PrEPS http www pressurebiosciences com pub application notes preparations html 1 Isolation of Functional Mitochondria from Rat Skeleta2. atteries Insert the charged battery into the SG3 Driver These batteries can be recharged at any time and do not have to be fully discharged before recharging HOW TO USE The SHREDDER SG3 Place approximately 10 to 300 mg of sample into the Ram side of an appropriate Shredder Tube Using the Shredder Tube Tool insert a serrated Shredder Ram with a twisting motion so that the sam ple is pressed between the serrated surface and the Lysis Disk Determine the method Shredder Tube type or re agent kit appropriate for your specific application by consulting the Table on Page 14 of this manual or by reviewing Application Notes and PrEPs on the PBI website The Shredder SG3 and Shredder Tubes are compati ble with a wide variety of sample preparation kits and reagents such as Trizol for RNA extraction as well as kits sold by PBI for specialized sample preparation applications http www pressurebiosciences com pub application notes preparations html Add 200 700 uL of appropriate extraction reagent to the Cap side of the tube The total volume of sample and buffer during shredding should not exceed 0 0 8 mL This prevents buffer from being forced into the threads of the Cap where it might be lost during uncapping
3. cess Further the FSL eliminates the need for the operator to exert force for long periods when processing one or more sam ples To recover any residual sample gently push the Shredder Ram with the Shredder Tube Tool until the Ram presses on the Lysis Disk Aspirate the re maining sample In some cases it may be neces sary to remove particulates from the homogenate by brief centrifugation prior to downstream processing Shred as long as necessary until all or most of the sample has passed through the Lysis Disk to the up per chamber of the Shredder Tube It may be neces sary to shred for several seconds using a lower lever setting before completing shredding on a higher set ting Note If PCT treatment will follow shredding the final volume of sample and reagents in the tube must be adjusted to 1 4 0 1 mL prior to placing the sample into the Barocycler Heat Generation While Shredding Many samples including most kinds of tissue sam ples require only 20 30 seconds of shredding While all shredding will produce some heat due to friction this short processing time will not signifi cantly heat the sample However harder samples like tree bark and seeds may need to be shredded for a longer time to achieve maximum homogenization In these cases more heat will be generated To minimize heat gen eration during longer shredding protocols the Stainless steel SHREDDER Base can be pre chilled by placing it into a refrige
4. eds or rice forcing the lever down to position 3 when shredding a sample larger than 25 mg or samples of soft tissue greater than 100 mg may damage the Lysis Disk It is best to shred briefly at position 2 until most of the sample is shredded Then proceed to position 3 to complete the shredding p Cap the Shredder Tube using a Shredder Screw Cap with the Shredder Tube Tool Place the filled Shredder Tube into the SHREDDER Base Ram side down and twist to set the Ram into the holder in the SHREDDER Base When the Tube is seated securely place the SG3 Driver onto the Cap and briefly turn the driver on in order to seat the Driver bit into the crenellations of the Cap 10 SHREDDER APPLICATIONS REQUIRING PRESSURE CYCLING TECHNOLOGY PCT PCT is not require for all shredder preparations However some preparations are enhanced when shredding is followed by PCT Please refer to Page 15 or the PBI website under Application Notes and Preparations PrEPs for spe cific examples of PCT enhanced extraction http www pressurebiosciences com pub application notes preparations html To obtain more information about a Barocycler for high pressure extractions please refer to the PBI website www pressurebiosciences com or ask your Sales Representative how PCT may enhance your particular application Figure 3 A PBI Barocycler NEP2320 7 PREPARING The SHREDDER SG3 First fully charge one of the two lithium ion b
5. hite Protein from Whole FT S00 MS 30 60 Rice Grains coral or seconds white Protein from Porcine FT 500 MS 40 60 Cartilage coral or seconds white 14 SAFETY INFORMATION Standard laboratory safety practices should be em ployed when working with chemicals to minimize po tential exposure to hazardous materials PRODUCT USE LIMITATIONS The SHREDDER SG3 is intended for Research Purposes Only No claim or representation is in tended to provide information for the diagnosis pre vention or treatment of a disease PRODUCT WARRANTY AND SATISFACTION GUARANTEE PBI warrants to the original purchaser only that all The SHREDDER SG3 parts will be free from defects in material and workmanship for a period of ninety 90 days from the date of purchase Sellers sole obliga tion and your exclusive remedy under this limited war ranty and to the extent permitted by law any warranty or condition implied by law shall be the repair or re placement of parts without charge which are defec tive in material or workmanship and which have not been misused carelessly handled or repaired by per sons other than PBI or authorized service representa tive lf The SHREDDER SG3 does not meet your expecta tions please contact our Technical Service Depart ment PBI reserves the right to change alter or mod ify any product to enhance its performance and de sign TECHNICAL ASSISTANCE At PBI we pride ourselves on the qual
6. ity and availabil ity of our technical support Our Technical Service Department is staffed by experienced scientists with 3 extensive practical and theoretical expertise in sample preparation proteomics molecular biology and the use of PBI products If you have any questions or ex perience any difficulties regarding The SHREDDER SG3 or other PBI products or services please do not hesitate to contact a Technical Service Representa tive at technicalsupport ressureblosciences com INTRODUCTION The SHREDDER SG3 is a low shear mechanical ho mogenization system for use with tough fibrous and other difficult to disrupt tissues and organisms The SHREDDER SG3 uses a variety of Shredder Tubes to directly and rapidly grind a biological sample providing easy handling and reducing sample contamination This low shear mechanical disruption process when combined with selected buffers can provide effective extraction of proteins DNA RNA lipids and small molecules from tissues and organisms Additional ex traction efficiency can be achieved when this process is combined with pressure cycling technology PCT resulting in further improvement of sample lysis and extraction of the desired analytes The SHREDDER SG3 features a three position force setting lever FSL Otherwise it is similar in function to PBI s The PCT Shredder The FSL enables the op erator to select and apply reproducible force to the sample during the shredding pro
7. l Muscle Using The PCT Shredder with an Optional Pressure Cycling Technology PCT Proc essing Step Isolation of Mitochondria from Rat Lung Using The PCT Shredder and Pressure Cycling Technology PCT Proteolysis Proteinase K PrEP Efficient and Rapid Extraction of Genomic DNA from Skeletal Muscle using The PCT Shredder and Pressure enhanced Proteolysis with Proteinase K Increased Protein Yields from Coniferous Plants Using The PCT Shredder and Pressure Cycling Technology PCT Improved Protein Recovery from Muscle Tissue Using The PCT Shredder and Pressure Cycling Technology PCT Efficient Single Tube Sample Disruption and Extraction Improved DNA Recovery from Spinach Leaves Us ing The PCT Shredder and Pressure Cycling Tech nology PCT PROTOCOLS for The SHREDDER SG3 The following Table lists some applications and gen eral conditions for The SHREDDER SG3 For com plete protocols consult the Application Notes and PrEPs under Shredder Applications Some General Conditions for The SHREDDER SG3 Application Shredder Shred Lever Tube Time Position DNA Extraction FT 500 S 20 30 2 from Spinach coral or seconds Leaves white DNA Extraction FT 500 MS 30 40 2 from Whole Apple coral or seconds Seeds white Protein Extraction FT 500 S 20 30 from Skin or Mus coral or seconds cle Tissue white RNA from Muscle FT 500 S 20 30 Tissue coral or seconds end RNA from Rodent FT 500 MS 20 30 Lung Tissue coral or seconds w
8. rator or a 20 C freezer Note Shredding non stop for 2 minutes is not recommended as it may result in decreased pro tein recovery due to heat induced protein aggre gation Removing the Processed Homogenate To remove the processed homogenate use the Shredder Tube Tool to open the Shredder Tube by removing the Shredder Cap Transfer the sample to a micro centrifuge tube or to another appropriate container 12 HARDWARE SUPPLIED with The SHREDDER SG3 a One Heavy Duty High Torque 1B SG3 Driver with a Convertible Handle B SG3 Base with a 3 position Force Setting Lever FSL fer Battery Charger Two Lithium lon Batteries os One Shredder Tube Tool One Assortment Package of FT500 SV Shredder Tubes Figure 1 The SHREDDER SG3 Kit Components Shown in Case Batteries Not Shown Figure 2 The SHREDDER SG3 Assembled for Shredding VARIATIONS AND SPECIFICATIONS OF SHREDDER TUBES PCT Tube Type Color Compatible PCT Shredder White N PULSE Tubes FT500 S Shredder Tubes Coral N FT500 PS Shredder Tubes Coral Y FT500 PMS PBI offers a variety of Shredder Tubes for various ap plications Only white Shredder Tubes are approved for use with high pressure sample preparation PCT Shredder White Yes Yes PULSE Tubes FT500 MS i While pressing down on The SHREDDER SG3 Driver with one hand set the lever into the appropriate posi tion 1 Low 2 Med or 3 High for the sample For hard samples such as se
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