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1. of accepting lots with given concentration levels It also plots the probabilities in graph form for easy interpretation Specific data can be saved for documentation and future analyses 2 The following is a helpful reference for use in designing a sampling plan Remund K M Dixon D A Wright D L Holden L R Statistical considerations in seed purity testing for transgenic traits Seed Science Research June 2001 Vol 11 No 2 pp 101 119 3 To select the appropriate sample size determine the purity standard and the QuickStix Kit for QuickScan Roundup Ready Bulk Soybeans Page 2 of 5 Choose grinding method Soybean Common Extraction Grams of beans x 5 mL water For example 100 x 0 15 I5 g I5 gx 5 75 mL water Extract in tap water z Transfer extract to cup about 3 pipettefuls Rey 02 01 13 degree of confidence required Confidence level means the statistical probability that the true Roundup Ready level in the lot is below the selected purity standard Table 1 provides a guideline for determining the number of 400 bean sub samples that are necessary to provide effective screening for different GM concentrations at a 95 confidence level Table 1 Number of 400 soybean subsamples required for testing Confidence Roundup Ready Level Soybean Screening Level For other sampling scenarios or different screening or confidence levels refer to the USDA GIPSA Excel spr2. ENVIRQLOGIX Highlights e Results in 5 minutes or less e Both bulk soybean kits utilize Common Extraction Roundup Ready LibertyLink e Available as 100 strip kit in bulk packaging or in QuickCombs Contents of Kit e 100 QuickStix Strips packed in two motsture resistant canisters e 100 transfer pipettes e 100 three ounce cups Items Not Provided e Blender for sample prep 1 Oster Sunbeam blender model 4094 with 4 oz polystyrene blender jar ice crusher blade gasket and blender base or 2 Waring blender model 3IBL91 or equivalent with glass Mason jars and jar adapter Eberbach E8495 along with protective cover 3 BUNN coffee grinder industrial style grinder set on AutoDrip setting e Graduated cylinder e Tap water e Protective cover for blender jar while grinding e QuickScan System optional for quantitative results Rey 02 01 13 QuickStix Kit for QuickScan Roundup Ready Bulk Soybeans Catalog Number AQ 010 BGB Intended Use This EnviroLogix QuickStix Kit for QuickScan Roundup Ready Bulk Soybeans is designed to extract and detect the presence of CP4 EPSPS protein at the levels typically expressed in Roundup Ready soybeans The sensitivity of these QuickStix Strips is 0 25 Roundup Ready soybean i e one soybean in 400 How the Test Works In order to detect the CP4 EPSPS protein expressed by Roundup Ready soybeans the sample must first be extracted t
3. Inc EnviroLogix warrants the products sold hereunder the Products against defects in materials and workmanship when used in accordance with the applicable instructions for a period not to extend beyond a product s printed expiration date If the Products do not conform to this Limited Warranty and the customer notifies EnviroLogix in writing of such defects during the warranty period including an offer by the customer to return the Products to EnviroLogix for evaluation EnviroLogix will repair or replace at its option any product or part thereof that proves defective in materials or workmanship within the warranty period NEITHER ENVIROLOGIX NOR MONSANTO MAKES ANY OTHER WARRANTIES EXPRESS OR IMPLIED INCLUDING BUT NOT LIMITED TO ANY IMPLIED WARRANTIES OF MERCHANTABILITY OR FITNESS FOR A PARTICULAR PURPOSE The warranty provided herein and the data specifications and descriptions of EnviroLogix products appearing in EnviroLogix published catalogues and product literature are EnviroLogix sole representations concerning the Products and warranty No other statements or representations written or oral by EnviroLogix employees agents or representatives except written statements signed by a duly authorized officer of EnviroLogix Inc are authorized they should not be relied upon by the customer and are not a part of the contract of sale or of this warranty EnviroLogix does not warrant against damages or defects arising in
4. eadsheet described under Step 1 above or call EnviroLogix Technical Support for assistance Step 2 Determine Sub sample Weight Jar Size and Grind Times 1 Determine average weight of individual grain to be tested weigh 100 beans divide by 100 2 Calculate the weight of the number of grains to be tested Number of grains X Average Weight Grain Table 2 lists the guidelines for jar size and grinding time according to sample weight 3 Choose an appropriate jar size and grind time based on the type of blender available for sub sample preparation see Table 2 Oster Sunbeam Blender with ice crusher blade is recommended over the Waring Blender for its bean grinding efficiency Note that bean grind time is longer and requires additional steps when using a Waring Blender Table 2 Soybeans Grind Time of Beans Sub sample on High approximate Blender Type weight g Jar size 0Z Speed Sunbeam 60 seconds 60 seconds For best results blend beans for 2 of total time remove the jar and shake to redistribute larger particles replace and resume grinding 20 seconds 4 Choose an appropriate jar size for the sample based upon Table 2 above Step 3 Prepare the Sample 1 Weigh sample into the appropriate vessel 2 Put protective cover over glass jars 3 Grind sample on high speed until all whole beans are finely ground 4 Add the volume of tap water calculated by the formula left 5 Cap and shake jar vigorously
5. kStix Kit for QuickScan Roundup Ready Bulk Soybeans Page 4 of 5 e The assay has been optimized to be used with the protocol provided in the kit Deviation from this protocol may invalidate the results of the test e The results generated through the proper use of this diagnostic tool reflect the condition of the working sample directly tested Extrapolation as to the condition of the originating lot from which the working sample was derived should be based on sound sampling procedures and statistical calculations which address random sampling effects non random seed lot sampling effects and assay system uncertainty A negative result obtained when properly testing the working sample does not necessarily mean the originating lot is entirely negative for the analyte or protein in question e Protect all components from hot or cold extremes of temperature when not in use e CAUTION Tightly closed containers of soy extract if left sitting for several hours may ferment and cause the lid or container to burst Dispose of extract when testing is complete Rey 02 01 13 QuickStix Kit for QuickScan Roundup Ready Bulk Soybeans Page 5of5 For Technical Support Contact Us At EnviroLogix 500 Riverside Industrial Parkway Portland ME 04103 1486 USA Tel 207 797 0300 Toll Free 866 408 4597 Fax 207 797 7533 e mail info envirologix com website www envirologix com LIMITED WARRANTY EnviroLogix
6. logy LLC EnviroLogix the EnviroLogix logo QuickStix QuickScan and Common Extraction are trademarks of EnviroLogix Inc EnviroLogix 2013 Rey 02 01 13
7. o solubilize the protein Each QuickStix Strip has an absorbent pad at each end The protective tape with the arrow indicates the end of the strip to insert into the reaction cup The sample will travel up the membrane strip and be absorbed into the larger pad at the top of the strip The portion of the strip between the protective tape and the absorbent pad at the top of the strip is used to view the reactions as described under Interpreting the Results Results may then be scanned and interpreted quantitatively with the EnviroLogix QuickScan System Please avoid bending the strips Sample Preparation Step 1 Determine Number and Size of Sub samples 1 Collect a composite sample according to USDA GIPSA instructions found in the following reference documents http archive gipsa usda gov reference library handbooks grain insp grbook1 bk1 pdf USDA Grain Inspection Handbook Book 1 Grain Sampling http archive gipsa usda gov biotech sample2 htm Guidance document entitled Sampling for the Detection of Biotech Grains http archive gipsa usda gov biotech sample1 htm Practical Application of Sampling for the Detection of Biotech Grains http archive gipsa usda gov biotech samplingplan1 xls This website provides a simple to use Sample Planner 29K Excel Spreadsheet The planner allows you to enter different assumptions in terms of sample size number of samples acceptable quality level and to determine the probability
8. shipping or handling or out of accident or improper or abnormal use of the Products against defects in products or components not manufactured by EnviroLogix or against damages resulting from such non EnviroLogix made products or components EnviroLogix passes on to customer the warranty it received if any from the maker thereof of such non EnviroLogix made products or components This warranty also does not apply to Products to which changes or modifications have been made or attempted by persons other than pursuant to written authorization by EnviroLogix THIS WARRANTY IS EXCLUSIVE The sole and exclusive obligation of EnviroLogix shall be to repair or replace the defective Products in the manner and for the period provided above EnviroLogix shall not have any other obligation with respect to the Products or any part thereof whether based on contract tort strict liability or otherwise Under no circumstances whether based on this Limited Warranty or otherwise shall EnviroLogix be liable for incidental special or consequential damages This Limited Warranty states the entire obligation of EnviroLogix with respect to the Products If any part of this Limited Warranty is determined to be void or illegal the remainder shall remain in full force and effect License EnviroLogix has developed this kit using proprietary reagents as well as reagents licensed from Monsanto Company Roundup Ready is a registered trademark of Monsanto Techno
9. tape NOTE Use extreme caution to prevent sample to sample cross contamination with grain fluids or disposables Interpreting the Results Development of the Control Line within 5 minutes indicates that the strip has functioned properly Any strip that does not develop a Control Line should be discarded and the sample re tested using another strip Results are scanned and interpreted quantitatively with the QuickScan System Place QuickStix Strip into the carrier slide in and press Read Test on the screen QuickScan will return a result as GMO or lt LOD less than the Limit of Detection Please consult the QuickScan User Manual for details Kit Storage QuickStix can be stored at room temperature or refrigerated for a longer shelf life Note the shelf life on the kit box for each storage temperature The kit may be used in field applications however prolonged exposure to high temperatures may adversely affect the test results Do not open the desiccated canister until ready to use the test strips Precautions and Notes e This kit is designed to be read visually as a screen for presence or absence and is also designed to be quantitative when used with the QuickScan System e This product is currently not applicable for use in any other crop or in leaf or individual seed testing e As with all tests it is recommended that results be confirmed by an alternate method when necessary Rey 02 01 13 Quic
10. until the entire sample is wet 20 30 seconds QuickStix Kit for QuickScan Roundup Ready Bulk Soybeans Page 3 of 5 Strip must develop a Control Line to be valid cut where indicated and read in QuickScan System depending on the number of grains Sample will begin to settle immediately and liquid can be drawn off at that time Avoid pulling up particles with the transfer pipette 6 Transfer 12 mL of the liquid extract from above the settled soybean solids into the three ounce sample cup The level of extract should be above the bottom of the arrow tape yet below the top of the arrow on the bottom portion of the strip 7 To prevent cross contamination thoroughly clean blender parts and jars of dust and residue prior to preparation of a second sample Use a new transfer pipette and cup for each sample How to Run the QuickStix Strip Test 1 Allow refrigerated canisters to come to room temperature before opening Remove the QuickStix Strips to be used Avoid bending the strips Reseal the canister immediately 2 Place the strip into the three ounce cup containing 12 mL of the liquid soybean extract The sample will travel up the strip 3 Allow the strip to develop for 5 minutes before making final assay interpretations Positive sample results may become obvious much more quickly 4 To retain the strip or for use in the QuickScan System cut off and discard the bottom section of the strip covered by the arrow
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