DNA/RNA Extraction Kit
Contents
1. taco Research Use Only DNA RNA Extraction Kit User Manual Manufacturer GeneReach Biotechnology Corporation TEL 886 4 2463 9869 Email sales 9 genereach com No 19 Keyuan 2 Rd Central Taiwan Science Park Taichung City 407 Taiwan Website www tacomag com 2011 10 taco DNA RNA Extraction Kit Content SVIMDOISsissssscscseadessescotencdoneceoncecssvebasessonsesesSvecdessdvosveceseosesessesseseosemesees 1 Kit COMPOMNeMt sis iscdcssccscscosicsstesseatescosscscdasssscessentessbecseatessonssvvdecssseese 2 A Reagents ue tutte ete tret ii ed eme ia 2 B Plate amp Sleeve For one time use only suus 3 SIZE 3 Materials and Equipments Required but Not Provided 4 Hore reo Oo PR ai 5 Intended ae 5 Important NOS sce 6 Nucleic Acid Extraction Procedure eere 7 A Use of taco Sticker m e o cee en Edo eR he 7 B Protocol ede 8 Product Limitations eeeee eee eee eese sees sienten etas tn ssns sensn 12 Troubleshooting ecee eere oa eoe oae aao ep rb so eroe reor o ee ra eseese era oea 13 EVIL 17 Sample Preparation essent eene rennen 17 taco DNA RNA Extraction Kit Appendix TM ES 18 A Storage of Nucleic Acid 18 B Quantification of Nucleic Acid sess 18 C2. Appendix I Sample Preparation B Animal Tissue ethanol preserved shrimp muscle i li lii lv Grind the tissue 20 mg with 500 ul Lysis Buffer in 1 5 ml micro centrifuge tube with disposable grinder Centrifuge at 12000 rpm for 5 minutes to spin down the debris Transfer 400 ul of the supernatant and 200 ul 95 ethanol to column 1 7 of 96 Well Extraction Plate Follow Table 1 from Step 2 to 7 for loading reagents The above sample preparation method is recommended for general muscle tissues which contain high volume of protein other sample types must be validated by users 17 taco DNA RNA Extraction Kit Appendix II A Storage of Nucleic Acid Extracted Nucleic Acid should be stored at 80 C B Quantification of Nucleic Acid The concentration of nucleic acid should be determined by measuring the absorbance at 260 nm in a spectrophotometer Use Hluting Buffer as the blank to calibrate the spectrophotometer If the purified nucleic acid needs to be diluted before the quantification the Eluting Buffer also has to be diluted before use Also the same dilution factor needs to be applied for calculation Collect the absorbance reading of purified nucleic acid at 260 nm and 280 nm The reading should be located between 0 1 and 1 0 An absorbance of 1 unit at 260 nm corresponds to 50 ug of nucleic acid per milliliter The ratio between the absorbance values at 260 nm and 280 nm gives an estimation
3. Nucleic Acid Automatic Extraction System taco Step pipette optional Disposable gloves Micro centrifuge tubes Micropipette and Filter tips p1000 p200 e 95 ethanol Phosphate buffer saline PBS taco DNA RNA Extraction Kit Introduction The taco DNA RNA Extraction Kit is designed for taco Nucleic Acid Automatic Extraction System Based on the magnetic separation technology homogenized sample cells are lysed and nucleic acid is captured by silica coated magnetic beads Washing Buffer is then applied to remove impurities and Eluting Buffer to recover nucleic acid from magnetic beads following serial washing steps This kit can extract viral DNA and RNA from shrimp muscle Other sample types must be validated by users Note For research use only Not intended for any animal or human therapeutic or diagnostic use Intended Use The taco DNA RNA Extraction Kit is intended to be used for extracting viral DNA and RNA from various sample types such as shrimp tissue The taco DNA RNA Extraction Kit has to be used with the taco Nucleic Acid Automatic Extraction System This product is intended to be used by professional users such as well trained laboratory technicians who are familiar with molecular biology techniques taco DNA RNA Extraction Kit Important Notes After receiving the kit please check the kit components for any damage Contact GeneReach Biotechnology Corporation
4. Purity of Nucleic Acd e entiere etena aii a eais 19 taco DNA RNA Extraction Kit Symbols Date of manufacturing Manufacturer LOT Lot number Catalogue number Do not reuse amp E FEL taco DNA RNA Extraction Kit Kit Components A Reagents taco DNA RNA Extraction Kit Cat No atc d rna Number of reactions 320 Reagent Name Volume Quantity Magnetic Bead 18 ml 1 bottle Lysis Buffer 1 bottle Washing Buffer A 2 bottles Washing Buffer B 2 bottles Eluting Buffer bottle User Manual 1 copy Treat all reagents as potential irritants Add 135 ml 95 ethanol to Washing Buffer A before use Mark the bottle label after adding ethanol Add 230 ml 95 ethanol to Washing Buffer B before use Mark the bottle label after adding ethanol taco DNA RNA Extraction Kit B Plate amp Sleeve For one time use only Product Name Amount pcs Cat No 96 Well Extraction Plate Mixing Sleeve taco Sticker Do not reuse the Plate amp Sleeve Storage All reagents should be sealed tightly in cool and dry place at room temperature The expiration date of the kit and each component are stated on the label of each item Do not use any reagent of the kit beyond the expiration date Users should check the expiration date before use as it could affect the accuracy of the result taco DNA RNA Extraction Kit Materials and Equipments Required but Not Provided taco
5. environment is under room temperature 16 30 C 14 h Use non recommended extraction instrument taco DNA RNA Extraction Kit Comments and suggestions Using non recommended instrument may influence the performance of taco DNA RNA Extraction Kit We strongly recommend user to apply DNA RNA Extraction Kit on taco Poor DNA RNA performance in downstream applications a Low volume of extracted DNA RNA after the extraction is finished b Insufficient wm DNA RNA is used in downstream application c Excess DNA RNA used in downstream application Repeat the extraction procedure with new sample by using 100 ul Eluting Buffer Quantify the extracted DNA RNA by spectrophotometer of the absorbance at 260 nm See Quantification of Nucleic Acid Appendix IT Excess DNA RNA can inhibit some enzymatic reactions Quantify the extracted DNA RNA by spectrophotometer of the absorbance at 260 nm See Quantification of Nucleic Acid Appendix IT 15 Low A 260 A 280 ratio a Absorbance reading at 320 nm was not subtracted from the absorbance readings at 260 nm and 280 nm taco DNA RNA Extraction Kit Comments and suggestions To correct for the presence of Magnetic Bead particles in the eluted solution an absorbance reading at 320 nm should be taken and subtracted from the absorbance readings obtained at 260 nm and 280 nm 16 taco DNA RNA Extraction Kit
6. of nucleic acid purity See Purity of Nucleic Acid Carryover of Magnetic Bead may affect the A26 reading but should not affect the performance of nucleic acid in downstream applications Concentration of nucleic acid sample 50 ug ml x A2590 A320 x dilution factor Total amount of nucleic acid concentration x volume of sample in milliliters 18 taco DNA RNA Extraction Kit C Purity of Nucleic Acid Purity is determined by calculating the ratio of absorbance at 260 nm to absorbance at 280 nm with a background correction at 320 nm i e Ax69 A320 A2go A320 A subtracted absorbance reading at 320 nm is to correct the presence of Magnetic Bead particles in the eluted solution An A o Ago ratio of 1 6 2 0 is indicative of highly purified nucleic acid 2010 GeneReach Biotechnology Corporation All rights reserved For research use only Not intended for any animal or human therapeutic or diagnostic use 19
7. or your local distributor if reagent bottles are damaged Do not use damaged kit as it could affect the accuracy of the result Pipette tips are all for one time use only Repeated usage will lead to cross contamination When working with chemicals please always wear a suitable lab coat disposable gloves and protective goggles Discard gloves if they are contaminated Do not combine components with different batch Avoid microbial contamination of the reagents To minimize the risk of infections from potentially infectious material we recommend working under a laminar hood until the samples are lysed This kit should only be used by trained personnel Disposal of waste must be compliant with local laws taco DNA RNA Extraction Kit Nucleic Acid Extraction Procedure A Use of taco Sticker For your convenience you may put the taco Sticker on top of reagent bottles and on the rim of 96 Well Extraction Plate to avoid human error a taco Sticker Plate Sticker Apply the Sticker on the rim of 96 Well Extraction Plate LB wam wa SEL wan we OE Bottle Sticker Apply the Sticker on top of each reagent bottle b Abbreviation Definition LB Lysis Buffer M Magnetic Bead WA Washing Buffer A WAM Washing Buffer A Magnetic Bead WB Washing Buffer B E Eluting Buffer taco DNA RNA Extraction Kit B Protocol a Load reagents into 96 Well Extraction Plate according to Table 1 at the room tempera
8. p the Hook of Mixing Sleeve to fasten the mortise See the illustration below f Press the Door button of taco to close the door and press Start button g After the extraction procedure discard the Mixing Sleeves first taco DNA RNA Extraction Kit h Take out the 96 Well Extraction Plate then press Reset button i Transfer the nucleic acid from column 6 and or 12 to the new micro centrifuge tubes for further use See Purity of Nucleic Acid Appendix II j It is strongly recommended to use freshly extracted nucleic acid for downstream applications such as amplification Otherwise the extracted nucleic acid should be kept at 80 C for long term storage See Storage of Nucleic Acid Appendix II Note Any deviation from the instruction may lead to a low recovery rate of the nucleic acid extract taco DNA RNA Extraction Kit Product Limitations The system performance has been validated by using infected shrimp muscle for viral nucleic acid isolation The user is responsible for validating the performance of the taco DNA RNA Extraction Kit for other particular use The kit and plastic parts are not intended for any therapeutic or diagnostic purposes for animals or humans 12 Troubleshooting Low DNA RNA yield a Magnetic Bead was not resuspended completely b Washing Buffer A and B did not contain ethanol taco DNA RNA Extraction Kit Comment
9. s and suggestions Before starting the procedure ensure that Magnetic Bead is fully resuspended Vortex for at least 5 seconds before first use and perform mild agitation before subsequent uses Ensure the correct volume of ethanol is added to Washing Buffer A and B tightly seal the reagent bottles to prevent ethanol from evaporating Repeat the extraction procedure with proper reagent is necessary when the ethanol was not added to Wash Buffer A and Wash Buffer B before use For the proper procedure of extraction please see Protocol 13 c Reagents were loaded in wrong order d Poor sample quality e Incorrect sample volume f Mixing Sleeve was not installed g Inappropriate operation environment taco DNA RNA Extraction Kit Comments and suggestions Restart the loading procedure with a new 96 Well Extraction Plate Ensure that all reagents were loaded on the well in the correct order Repeat the extraction procedure with new samples Using fresh sample for extraction is recommended Poor sample quality may influence test result The kit performance would be affected if user did not use the right volume of sample User should optimize the sample quantity when dealing with different sample types Contact your local distributor or GeneReach Biotechnology Corporation for assistance Operation temperature could affect the recovery rate Please ensure the operation
10. ture 16 30 C for the best performance Table 1 Loading Reagent Step Reagents Add 200 pl 95 ethanol and 500 ul Lysis Buffer to column 1 7 2 Add 750 pl Washing Buffer A to column 2 8 3 Add 750 ul Washing Buffer A to column 3 9 4 Add 750 pl Washing Buffer B to column 4 10 5 Add 750 pl Washing Buffer B to column 5 11 6 Add 50 ul Eluting Buffer to column 6 12 7 Add 50 pl Magnetic Bead to column 2 8 Ensure that 135 ml 95 ethanol has been added to Washing Buffer A before the first time use Ensure that 230 ml 95 ethanol has been added to Washing Buffer B before the first time use 3 Magnetic Bead must be resuspended before aliquoting taco DNA RNA Extraction Kit b Grind the tissue 40 mg with 450 ul PBS in a 1 5 ml micro centrifuge tube with disposable grinder Centrifuge at 12000 rpm for 5 minutes to spin down the debris For ethanol preserved sample please see Sample Preparation Appendix D c Transfer 200 ul of the supernatant to column 1 7 of 96 Well Extraction Plate d Open the door of taco and install the 96 Well Extraction Plate with reagents and samples Push 96 Well Extraction Plate completely into the bottom of plate holder Ensure the cut site is located on the top right F cut site IEELEPELEBELRDBLILIS 1000090009099 taco DNA RNA Extraction Kit e Install the Mixing Sleeve and lift u
Download Pdf Manuals
Related Search