Total DNA Extraction Kit
Contents
1. TS 96 Well Extraction Plate Mixing Sleeve PB atcp s taco Sticker Do not reuse the Plate amp Sleeve taco Total DNA Extraction Kit Storage amp Shipping All reagents should be stored well sealed and kept dry at room temperature 2344 C up to the expiration date labeled on the box Deliver all reagents at room temperature if necessary Expiration dates are stated on the box and on each single component of the kit Do not use any component of the kit beyond the expiration date Any deviation from the instruction could influence the kit performance and must be validated by the users Table 1 The stability of taco Total DNA Extraction Kit Stability Shelf life year Open vial stability 30 days Concentration of Extracted DNA The concentration of DNA depends on the sample type number of nucleated cells in the sample and the protocol used for isolation of DNA Table 2 shows concentration obtained from different sample volumes and sample types taco Total DNA Extraction Kit Table 2 DNA concentration obtained from whole blood and buffy coat using taco Total DNA Extraction Kit Sample type Sample volume ul Concentration ng pl Whole blood 200 gt 40 Buffy coat 150 gt 80 The normal range of white blood cell WBC count from fresh samples is 5000 10000 cells ul Equipment and Reagents to Be Supplied by Users taco Nucleic Acid Automatic Extraction System taco Step pipette opti2. Product Use Limitations The system performance has been validated using human whole blood and buffy coat for the isolation of genomic DNA The kit is neither validated for use with bone marrow cultured cells nor for the isolation of total DNA from serum plasma or nor for the isolation of total RNA The user is responsible for validating the performance of the taco Total DNA Extraction Kit for any particular use To minimize the risk of a negative impact on the diagnostic results adequate controls for downstream applications must be used Any diagnostic results that are generated must be interpreted in conjunction with other clinical or laboratory findings 13 Trouble Shooting Low DNA yield a Magnetic Bead was not completely resuspended b Washing Buffer B did not contain ethanol c Reagents were loaded in wrong order taco Total DNA Extraction Kit Comments 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 B well seal the reagent bottle to prevent ethanol from evaporating Add 50 ml 95 ethanol to Washing Buffer B before use Repeat the extraction procedure with proper reagent Restart the loading procedure with a new 96 Well Extraction Plate Ensure that all reagents were loa
3. A Storage of DNA Extracted DNA should be stored at 2 8 C for 24 hours or at 20 C for longer storage B Quantification of DNA The concentration of DNA should be determined by measuring the absorbance at 260 nm A269 in a spectrophotometer Use Eluting Buffer as the blank to calibrate spectrophotometer If the purified DNA needs to be diluted before the quantification the Eluting Buffer also has to be diluted first and the same dilution factor needs to be applied for calculation Collect the absorbance reading of purified DNA at 260 nm and 280 nm The reading should fall between 0 1 and 1 0 to be accurate An absorbance of 1 unit at 260 corresponds to 50 ug of DNA per milliliter The ratio between the absorbance values at 260 nm and 280 nm gives an estimation of DNA purity See Purity of DNA Carryover of Magnetic Bead may affect the Ar reading but should not affect the performance of DNA in downstream applications Concentration of DNA sample 50 ug ml x A250 A320 X dilution factor Total amount of DNA purified concentration x volume of sample in milliliters 18 C Purity of DNA Purity is determined by calculating the ratio of corrected absorbance at 260 nm to corrected absorbance at 280 nm Le Ax69 A320 A2go A320 A subtracted absorbance reading at 320 nm is to correct the presence of Magnetic Bead particles in the eluted solution The purity of DNA has an A260 Ago ratio of 1 6 2 0 201
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5. age Contact GeneReach Biotechnology Corporation or your local distributor if the reagent bottles are damaged Do not use damaged kit components since their use may lead to poor kit performance Always change pipette tips between liquid transfers When working with chemicals always wear a suitable lab coat disposable gloves and protective goggles Discard gloves if they become contaminated Do not combine components of different kits Avoid microbial contamination of the kit reagents To minimize the risk of infections from potentially infectious material we recommend working under laminar air flow until the samples are lysed This kit should only be used by trained personnel Dispose of waste must be compliant to local laws taco Total DNA Extraction Kit Safety Information To avoid injuries when working with the kit component always wears proper clothing and accessories as recommended For more information please consult the appropriate Material Safety Data Sheet MSDS CAUTION DO NOT add bleach or acidic solutions directly to the sample preparation waste The Lysis Buffer and Washing Buffer A contain guanidine thiocyanate which can form highly reactive compounds when combined with bleach Should any spillage occur clean with suitable laboratory detergent and water Broken and leaky reagent buffers must be handled and discarded according to local safety regulations Do not use damaged reagent buffers or oth
6. antification of DNA Appendix II Excess DNA can inhibit some enzymatic reactions Quantify the extracted DNA by spectrophotometer of the absorbance at 260 nm See Quantification of DNA Appendix II A260 A289 ratio for extracted DNA is low a Absorbance reading at 320 nm was not subtracted from the absorbance readings at 260 nm and 280 nm To correct 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 Total DNA Extraction Kit Appendix I A Sample Preparation whole blood and buffy coat Add 200 ul of whole blood or 150 ul buffy coat sample into column 1 7 of 96 Well Extraction Plate contains 400 ul of Lysis Buffer B Preparation of Buffy Coat About 10 ml of whole blood in an EDTA vacutainer tube Prepare buffy coat by centrifuging the tube at 3000 x g for 10 minutes in room temperature After centrifugation 3 different layers are easy to distinguish the upper layer is plasma the intermediate layer is buffy coat containing concentrated leukocytes and the bottom layer contains concentrated erythrocytes Transfer the middle layer to a new tube carefully It may contain small amounts of plasma and concentrated erythrocytes aspirates off the plasma layer carefully may help harvesting the buffy coat 17 taco Total DNA Extraction Kit Appendix II
7. ations siscccecessseccsccssecessstvscstacscaseieiseeatccssvaseaash veces 13 Trouble SHO OMG a aaaea oraaa aaaeeeaa aea Oaea EEEE AEE ETRE 14 taco Total DNA Extraction Kit PODCTIOIX Toseta oaaae AE aa EERE 17 Ac Salnip le Prepa AON srei ea ELU e EEEa 17 Bo Preparatiomof Butiy COl ecoiiniis nonan ER NA 17 APPENAdIX aenean eR or WES EEEN EOAR a ERS 18 A Storage Of DNA cisssscsucacdcacecassssausatasees savedesuedsdansecatsistustbessssensces 18 B Quantification of DNA cissesesescsessscseessssessiasseesscouseevacsesscstucteeunes 18 Co Punt wor DNAS ee eaea tar OAE EEEE 19 ll Symbols E lt 0 m O j gt IBOLE taco Total DNA Extraction Kit Contains reagents for 64 reactions Expiration date in vitro diagnostic medical device Catalogue number Lot number Manufacturer Temperature limitation Date of manufacturing Do not reuse Consult the User Manual for use EU Authorized Representative Caution Not following the instruction may lead to poor kit performance or personnel injured taco Total DNA Extraction Kit Kit Components A Reagents taco Total DNA Extraction Kit Cat No atci dna Number of reactions Eluting Buffer 1 bottle es ee o Treat all reagents as potential irritants Add 50 ml 95 ethanol molecular biology grade to Washing Buffer B before use Mark the label of reagent bottle after the addition of ethanol B Plate amp Sleeve For single use
8. ded in the correct order and wells Repeat the extraction procedure with new sample 14 d Low leukocyte count in the whole blood sample e Poor sample quality f Mixing Sleeves not installed g Inappropriate operation environment h Use non recommended extraction instrument taco Total DNA Extraction Kit Comments and Suggestions Increase whole blood amount and keep the volume of buffy coat harvested constant Using fresh sample for extraction 1s recommended Poor blood quality may influence test result Contact your local distributor or GeneReach Biotechnology Corporation for assistance Operation temperature too high or low may lead to low yield of DNA Ensure the operation environment of taco Total DNA Extraction Kit to be performed only under room temperature 16 30 C User uses non recommended instrument may influence the performance of taco Total DNA Extraction Kit We recommend user to apply it on taco Poor DNA performance in downstream applications a Low volume of extracted DNA after the extraction is finished Repeat the extraction procedure with new sample using 100 ul Eluting Buffer 15 b Insufficient DNA used in downstream application c Excess DNA used in downstream application taco Total DNA Extraction Kit Comments and Suggestions Quantify the extracted DNA by spectrophotometer of the absorbance at 260 nm See Qu
9. er kit components since their use may lead to poor kit performance CAUTION Reagent buffer contains EtOH such as Washing Buffer B must be kept out of flame After the addition of EtOH to the Washing Buffer B ensure the bottles are kept away from flame since it could be highly flammable harmful and irritant taco Total DNA Extraction Kit Nucleic Acid Extraction Procedure A Use of taco Sticker For your convenience user may put the taco Sticker on top of reagent bottles and or 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 itp wam wa M E e wan w Sc Bottle Sticker Apply the Sticker on top of each reagent bottle LB M WA WA ow E 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 Total DNA Extraction Kit B Protocol a Load reagents into 96 Well Extraction Plate according to Table 3 below at the room temperature 16 30 C for the best kit performance Table 3 Loading reagent 5 Aas plang Batter o eoma 62 Ensure that 50 ml 95 ethanol has been added to Washing Buffer B before the first time use Magnetic Bead must be mixed until it s fully resuspended before each aliquot b Use micropipette to load homogenized samples into colum
10. n 1 and or 7 See Sample Preparation Appendix I taco Total DNA Extraction Kit c Open the door of taco and install 96 Well Extraction Plate with reagents and samples Push 96 Well Extraction Plate completely to the bottom of plate holder Ensure the cut site is located on the top right p cut site feb GB GD GB aD GD DED ENED ENED SOGSOSOSSSSsS ep Oo Oe Oe Re ep Oe OD Om OD w Gp Ge Go Ge Go Ge Ge Gp Ge GD GG 500000000008 S000060 00000S SD So Gp am Ge Gp We Gp ep am Op wp j SD SP Go Go Gp Gd GD Gp GP GP Ge Go 10 taco Total DNA Extraction Kit d Install Mixing Sleeve and lift up the Hook of Mixing Sleeve to tenon the mortise See the illustration below 11 taco Total DNA Extraction Kit e Press the door button of taco to close the door and press Start button f After extraction finished discard the Mixing Sleeves first g Take out the 96 Well Extraction Plate and press Reset button h Transfer the total DNA from column 6 and or 12 to new micro centrifuge tubes for use See Purity of DNA Appendix ID i It is strongly recommended to use the freshly extracted DNA for downstream applications such as amplification Otherwise the extracted DNA should be kept at 20 C for longer storage See Storage of DNA Appendix II Note Any deviation from the instruction may _ affect the kit performance 12 taco Total DNA Extraction Kit
11. onal Disposable gloves Micro centrifuge tubes Micropipette p1000 p200 Filter tips p1000 p200 95 ethanol molecular biology grade taco Total DNA Extraction Kit Introduction The taco Total DNA Extraction Kit is designed for taco Nucleic Acid Automatic Extraction System Based on the magnetic separation technology homogenized sample cells are lysed and nucleic acids are captured by silica coated magnetic beads Washing Buffer is then applied to remove impurities and Eluting Buffer to recover nucleic acids from magnetic beads following serial washing steps This kit can extract total DNA from whole blood and buffy coat Other sample types must be validated by users Intended Use The taco Total DNA Extraction Kit is intended to be used for extracting total DNA from human whole blood and buffy coat for subsequent in vitro diagnostic application The taco Total DNA Extraction Kit is designed 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 familiar with molecular biology techniques Any diagnostic result generated from the sample preparation procedure followed by any downstream diagnostic nucleic acid testing assay must be complemented by other clinical or laboratory finding taco Total DNA Extraction Kit Important Notes After receiving the kit check the kit components for any dam
12. taco CE Total DNA Extraction Kit User Manual Manufacturer GeneReach Biotechnology Corporation TEL 886 4 2463 9869 Email sales genereach com No 19 Keyuan 2 Rd Central Taiwan Science Park Taichung City 407 Taiwan Website www tacomag com EC REP Wellkang Ltd www CE marking eu 29 Harley St London W1G 9QR UK 2011 04 taco Total DNA Extraction Kit Content DP VMIDOIS scesccsccssevestccscsccsssssvesssesssestesssvestectusceseasbbesesesssesseaseeastecsuccstuesees 1 Kit COMPONE S kasetin bconckcersaseoeseeseudes sectoceteeneeesseaseoeseeeweees 2 PINS AOC aE a 2 DPBS ee SISSY Circe nas eet ihc ee a Uhh aan 2 Storage amp Shipping sos scesesscsessvsessscsssoveasecssswensvssvteesvssavicseteccatecsesveesvs 3 Concentration of Extracted DNA ssseessssssssssssooccccceccecsesesssssosssssoe 3 Equipment and Reagents to Be Supplied by Users s000 4 TRE OCU CTION wasacesccasstisecitatesaseccsruaatiate aeacsisdecitnaesasascarvaticteacatsisiecitens 5 Mtended USO ssi debs csceveseeues a Ea 5 D portant Notes ssssscccseceicsdiccssscevecstnstisssssueiivedsedessssesessuscessenssseecivesises 6 Safety Mmiormatio D esne i N 7 Nucleic Acid Extraction Procedure cssssssssssssssssssccccccssssees 8 A Use of taco STICKER sacaictastedocsssinsrsiaiudacdsantoustsaladeovaneretaeodeess 8 Pe eg RG 8 0 0 Were Teen rere tenes enter er Ot fms emne ero tanes nt ee ner er free Tene mere 9 Product Use Limit
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