PDS - BIORON GmbH
Contents
1. Calculate the amount of sample plus controls and pipette the whole amount of needed volume to an appropriate tube E g for 10 samples use 250 ul Magnetic Beads Vortex the tube of magnetic beads for 15 30 seconds to ensure that the particles are thoroughly a homogeneous suspension Place the tube with the magnetic beads in a magnetic separator in order to collect the beads to the side of the tube Wait approximately 30 60 seconds until the beads have been attracted to the magnet The supernatant has to be clear If not prolong the time until the supernatant is clear Discard the supernatant by using pipette with filter tip and then remove the tube from the separator Re suspend for each sample the magnetic bead pellet in 75 ul Ilsopropylalcohol 70 v v If using the whole amount of magnetic beads pipette the respectively amount of needed volume E g for 10 samples use 750 ul Isopropylalcohol 70 v v In case of using a repeater pipette in step 5 7 it is recommended to calculate one sample more E g for 10 samples use 825 ul prepared Magnetic Beads volume for 11 samples Store at 2 8 C For optimization of time this step can be done during sample lysis step 6 5 BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 14 of 19 PureMagic BIORON 6 4 Sample Preparation The sample2. 19 Table of 06012015IH BIORON PureMagic Content MTG OMS ite aE a can ats Gated E iota Va casa E E E acne t Mian E 4 Specification of the Kil icc ccvccrwsandcark ecbamneiencaccesereecewadedaeiecnbecennneoniedeccthacncctdneeharinaeeencars 5 Components of the Kit and Storage ccccccceeeeeeeeeeseceeeeeeeeeeessecaaeeeeeeeeeeeesesnaaeeeeeees 6 Description of the Method Principle eeeeeccceeceeeeeeeeeeeeeneeeeeeeeeeeeeeeeaaaaneeeeeeenenas 7 Extraction PIOWWCOl 5irscirncarncaints muren issn sns in ni iaa r ina i ian ii 7 Procedure for Automated DNA Purification for the KingFisher Flex n 13 Troubleshooting TipS sssseeeeesoeeeneeenteeererettrnntrsetrtrtrrnnntrsterttrtnnnreserrtntrnntnseerne nenna 18 BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net Page 3 of 19 PureMagic BIORON 1 Introduction The efficient isolation of DNA of a wide range of interesting specimen becomes more and more an important tool for testing of food and environmental safety DNA can be used in high sensitive and specific methods like PCR and real time PCR for the subsequent detection of pathogenic organisms and unacceptable additions in any kind of food The PureMagic Food Extraction Kit is designed for the isolation of high quality nucleic acids from a variety of raw and processed food samples This DNA is suitable for downstream methods
3. like PCR or sequencing Therefore the possibility of identification of animal components in food and feed is given Typical applications for PureMagic are species detection in food cosmetics and pathogen detection in feed and food The DNA binding magnetic particles chosen for extraction are optimized for DNA isolation and purification The high magnetic strength makes them applicable for both manual and automated DNA isolation e g KingFisher Flex Thermo Fisher Scientific BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 4 of 19 PureMagic BIORON 2 Specification of the Kit The PureMagic Extraction Kit is designed as a purification system for nearly all types of food and feed samples This system have been optimized for the isolation of DNA from canned and processed food e g salted colored and dried food for the subsequent detection of DNA of animals like pork in the products In addition DNA can be purified not only from milk and the co products such as milk powder cheese and yoghurt but also the lysis buffer was also tested for extraction of DNA from plant bacteria and cosmetics see list below Raw canned and highly processed products meat from cow turkey chicken camel horse pork fish Sausage Milk Cheese Yoghurt Milk Powder Milk Drinks Food animal origin Vegetable oils Margarine Break
4. 0 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 9 of 19 PureMagic BIORON 5 5 Sample Preparation The samples are lysed under denaturing conditions in the presence of Lysis Buffer and Proteinase K This kit works for following samples in which sample preparation is different for liquids and non liquids Species detection a Non Liquid Liquid Bacterial samples samples Analysis go to 5 5 1 go to 5 5 2 go to 5 5 3 All substances that can be pipetted by means of a microliter pipette are termed Liquids All others are Non Liquids and have to be processed with the Non Liquids procedure 5 5 1 Non Liquids and other sample material Weigh 50 mg 5 mg sample material into a 1 5 or 2 0 ml reaction tube e g by using scalpels or tooth pickers Be aware of cross contaminations gt Proceed with lysis step 5 6 5 5 2 Liquid samples Vortex the samples thoroughly Use an amount of 300 ul for all liquids samples gt Proceed with lysis step 5 6 5 5 3 Samples for Bacterial Analyse in liquids milk products Pipet 1 ml of the liquid into a 1 5 or 2 0 ml reaction tube and pellet bacteria by centrifugation for 10 minutes at 5 000 x g Discard the supernatant and use the pellet for DNA extraction gt Proceed with lysis step 5 6 Note Please avoid pipetting milk clots from milk samples into the reaction tube BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany
5. Dry the particles at 65 C for 3 minutes tube with open cap gt Proceed with step 5 9 5 9 DNA Elution Elution Buffer rehydrates the DNA so it will elute from the bead 1 Add 75 pl Elution Buffer to each sample and vortex 2 Incubate at 65 C for 5 minutes and 1 400 rpm on the thermomixer 3 Place the tube in the magnetic separator and leave in place for approx 30 60 seconds to collect the beads 4 Carefully transfer the eluate to a new reaction tube The DNA in the eluate is now ready to use It is recommended to apply the DNA immediately or after storage at 4 8 CT over night Note If a large amount of DNA is expected the volume of elution buffer can be increased BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 12 of 19 PureMagic BIORON 6 Procedure for Automated DNA Purification for the KingFisher Flex The following protocol is designed for general use with the KingFisher Flex instrument 96 Deep Well Head Automation is based on magnetic rods that move particles through the various purification phases including binding mixing washing and elution 6 1 Reagents Equipment and Consumables supplied by the User Reagents and Solutions e 70 lsopropylalcohol lsopropanol 2 Propanol v v e 100 Isopropylalcohol Isopropanol 2 Propanol v v Laboratory Material e Microliter pipettes a
6. Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 10 of 19 PureMagic BIORON 5 6 Sample Lysis 1 Prepare a mix of Lysis Buffer and Proteinase K For each sample 250 ul Lysis Buffer and 50 pl Proteinase K LP mix is used Mix well Add 300 ul of that LP mix to each sample tube Calculate a master mix for easier pipetting 2 Incubate the sample tubes for 1 hour at 55 C and 600 rpm on the thermomixer Note The sample should be completely lysed If necessary prolong the incubation time no visible particles left some turbidity is appropriate 3 Centrifuge the sample for 3 minutes at 16 000x g 4 Transfer the supernatant lysate into a new reaction tube gt Proceed with step 5 7 Note The presence of fat on top of the supernatant following centrifugation is normal Remove carefully the supernatant without touching the fat layer 5 7 DNA Binding 1 Add 350 pl 70 Isopropylalcohol v v to the lysate and mix well to get a homogeneous suspension 2 Add 75 ul of the prepared beads see 5 4 Mix beads by vortexing before adding them to the sample gt Optional Add the whole amount of prepared beads to the appropriate volume of 70 Isopropylalcohol v v E g for 10 samples use 750 ul prepared magnetic beads plus 3500 ul 70 Isopropylalcohol In case of using a repeater pipette it is recommended to calculate one sample more 3 Mix sample and incubate for 15 minutes on the
7. Press Start after removing each plate Press Stop after all plates are removed 6 7 Storage After finishing the extraction protocol the Elution Plate contains the extracted DNA Use the DNA directly or after storage at 4 8 C over night Storage the extracted DNA Either transfer the DNA into appropriate tubes or keep the eluated DNA in the plate and seal the plate Note If the extracted DNA contains carryover of magnetic particles transfer the DNA into a 1 5 ml reaction tube and centrifuge at maximum speed for 1 minute Transfer the clear supernatant contains the DNA into a new tube BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 17 of 19 BIORON 7 Troubleshooting Tips PureMagic Problem Cause Corrective action Incomplete lysis Reduce sample volume weight to 50 Prewarm elution buffer to 60 C Incomplete elution Drying of Wash Buffer may have been incomplete Prolong the time of collecting the beads Incomplete collection of steps binding washing and elution We magnetic beads recommend an incubation time up to 2 minutes Decrease the drying time Over drying of the magnetic Leaving the cap off of the reaction tube beads after washing step during the 65 C drying step for the indicated time to eliminate any residual alcohol Inefficient binding
8. PureMagic BIORON Instruction for Use PureMagic Food Extraction Kit KIT FOR THE PURIFICATION OF DNA FROM CANNED AND HIGH PROCESSED FOOD OR FROM OTHER PRODUCTS FOR THE SUBSEQUENT ANALYSIS IN REALTIME PCR OR OTHER MOLECULARBIOLOGICAL METHODS Product Size Catalog no PureMagic Food extraction 10 preps SBI87010 PureMagic Food extraction 50 preps BI87050 PureMagic Food extraction 100 preps BI87100 PureMagic Food extraction bulk BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 1 of 19 PureMagic BIORON Production of an individual bulk for automated applications on request Please store components immediately after delivery at following conditions Magnetic beads 2 8 C Proteinase K 18 22 C All remaining components should be stored 15 25 C Explanation of symbols used in labeling LOT Batch code REF Catalogue number Expiry date Temperature limitation Consult instructions for use Number of tests ma R 22 36 38 Harmful S 22 ial BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 2 of
9. cautions have to be followed Wearing of gloves as well as the application of filter tips is urgently recommended to minimize potential cross contamination For verification each examination should be carried out twice double examination e g prepare two DNA isolates from one sample In addition a negative extraction control without sample material and a positive extraction control with a known sample should be carried out in parallel with each DNA isolation Dispose unused reagents and waste in accordance with country federal state and local regulations Do not use the kit after the expiration date BIORON GmbH shall have no liability for any direct indirect consequential or incidental damages arising out of the use the results of use or the inability to use this product e amp lt eo amp amp BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 7 of 19 PureMagic BIORON 5 2 Reagents Equipment and Consumables supplied by the User Reagents and Solutions 70 Isopropylalcohol Isopropanol 2 Propanol v v 100 Isopropylalcohol Isopropanol 2 Propanol v v Laboratory Material e Microliter pipettes and sterile filter tips optional repeater pipette and PD Tips e 1 5 or 2 0 ml reaction tubes nuclease free all disposables are preferably low binding e Protective gloves Instruments e Vor
10. fast cereals and Muesli Food plant origin Jam Cookies cakes biscuits Jellies candies marsh mallows and chocolate products Pharmaceuticals Nutritional supplements like vitamin preparations Animal ingredi in e g cr make up sh Cosmetics ima i gredients in e g cr mes make up shower gel and body lotion Bacterial DNA from milk and other samples Enterobacter Listeria Staphylococcus Bacteria Streptococcus Clostridie E coli Baccilae Others Gelatine animal feeding stuff BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 5 of 19 BIORON 3 Components of the Kit and Storage PureMagic The delivery is carried out at RT so see storage condition as indicated on the bottle or tube PureMagic Food extraction Lysis Buffer Proteinase K Dilution Buffer for Proteinase K Wash Buffer Concentrate Elution Buffer Magnetic Beads RT Room temperature 15 10 preps 2x 1 5 ml 1x 11mg 1 x 600 ul 1x6ml 1x2ml 1 x 300 ul 25 C 50 preps 1x14ml 1x55 mg 1x3 ml 3x9ml 3x2ml 1x1 4ml 100 preps 2x14 ml 2x 55mg 2x3ml 6x9ml 6x2ml 2x1 4ml Colour code Storage at RT 18 22 C RT RT RT 2 8 C All PureMagic components have a guaranteed stability and the individual expiry dates are respectively indicated on the labels After reconstit
11. h sample 250 ul Lysis Buffer and 50 pl Proteinase K LP mix is used Mix well Add 300 ul of that LP mix to each sample tube Calculate a master mix for easier pipetting 2 Incubate the sample tubes for 1 hour at 55 C and 600 rpm on the thermomixer Note The sample should be completely lysed If necessary prolong the incubation time no visible particles left some turbidity is appropriate 5 Centrifuge the sample for 3 minutes at 16 000x g 6 Transfer the supernatant lysate into the according well of the lysate binding plate gt Proceed with step 6 6 Note The presence of fat on top of the supernatant following centrifugation is normal Remove carefully the supernatant without touching the fat layer 6 6 Preparation of Instrument and Plate Set Up For optimization of time this step can be done during sample lysis step 6 5 Note Before starting the purification process with the KingFisher Flex instrument please read carefully the user manual Resuspend Vortex the Magnetic Beads thoroughly directly before use Prefill the Binding Plate the Washing Plates and the Elution Plate as described below one well per sample Tip Plate Place the KingFisher Flex 96 Tip Comb for Deep Well Magnets on a Tip Plate Use one provided KingFisher Flex 96 KF Plate as Tip Plate These are identical Binding plate prepare a mix of magnetic beads see 6 3 5 1 and Isopropylalcohol 70 v v and mix well Add 425 ul of this mi
12. ient applications sample DNA Magnetic beads remaining in Perform one more magnetic separation step the eluate BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 19 of 19
13. nd sterile filter tips optional repeater pipette and PD Tips e 1 5 or 2 0 ml reaction tubes nuclease free all disposables are preferably low binding e Protective gloves e Microtiter Deep Well DW 96 Plate V bottom 100 1000 ul Product No 95040450 e KingFisher Flex 96 Tip Comb for Deep Well Magnets Product No 97002534 e KingFisher Flex 96 KF Plate Product No 97002540 Instruments e KingFisher Flex System 96 Deep Well Head Product No 5400630 e Rotator e Thermomixer at 65 C e Vortex mixer e Analytical Lab Scale e Tabletop centrifuge capable of 16 000 x g e Scalpel or disposables like tooth pickers for the food samples BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 13 of 19 PureMagic BIORON 6 2 Before starting If in Lysis Buffer a precipitate is formed dissolve it by incubating at 40 50 C till solution is clear Add 21 ml of 100 Isopropylalcohol to the Wash Buffer before using it for the first time Note After adding Isopropylalcohol Wash Buffer is stable for a minimum of 1 year at RT Proteinase K is supplied in a special formulated storage buffer After delivery we recommend storage at 18 20 C 6 3 Bead Preparation Note This step is strongly recommended for an optimal binding of the DNA to the beads for the best result For each sample 25 ul Magnetic Beads are used
14. reparation Note This step is strongly recommended for an optimal binding of the DNA to the beads for the best result 1 For each sample 25 wl Magnetic Beads are used Calculate the amount of sample plus controls and pipette the whole amount of needed volume to an appropriate tube E g for 10 samples use 250 ul Magnetic Beads 2 Vortex the tube of magnetic beads for 15 30 seconds to ensure that the particles are thoroughly a homogeneous suspension 3 Place the tube with the magnetic beads in a magnetic separator in order to collect the beads to the side of the tube Wait approximately 30 60 seconds until the beads have been attracted to the magnet The supernatant has to be clear If not prolong the time until the supernatant is clear 4 Discard the supernatant by using pipette with filter tip and then remove the tube from the separator 5 Re suspend for each sample the magnetic bead pellet in 75 ul Isopropylalcohol 70 v v If using the whole amount of magnetic beads pipette the respectively amount of needed volume E g for 10 samples use 750 ul Isopropylalcohol 70 v v In case of using a repeater pipette in step 5 7 it is recommended to calculate one sample more E g for 10 samples use 825 ul Magnetic Beads volume for 11 samples Store at 2 8 C For optimization of time this step can be done during sample lysis step 5 6 BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 572
15. rotator at 20 rpm or in the thermomixer at 1500 rpm and room temperature to allow the DNA binding to the beads 4 Place the sample in a magnetic separator in order to collect the beads to the side of the tube Wait approx 30 60 seconds until the beads have been attracted to the magnet 5 Discard the supernatant using pipette with filter tip and then remove the tube from the magnetic stand gt Proceed with step 5 8 Note Avoid losing the beads by carefully pipetting and contaminations by using fresh tips BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 11 of 19 PureMagic BIORON 5 8 Washing Step Contaminants are efficiently washed away using Wash Buffer while the DNA remains bound to the magnetic beads 1 Add 500 pl Wash Buffer to the tube and mix well by vortexing 2 Place the tube in a magnetic separator and leave in place for approx 30 60 seconds to collect the beads 3 Discard the supernatant by pipetting Remove as much of the liquid phase as possible and remove the tube from the stand 4 Add 1000 wl Wash Buffer to the tube and mix well by vortexing 5 Place the tube in a magnetic separator and leave in place for approx 30 60 seconds to collect the beads 6 Discard the supernatant by pipetting Remove as much of the liquid phase as possible after and remove the tube from the stand 7
16. s are lysed under denaturing conditions in the presence of Lysis Buffer and Proteinase K This kit works for following samples in which sample preparation is different for liquids and non liquids Species detection a Non Liquid Liquid Bacterial samples samples Analysis go to 6 4 1 go to 6 4 2 go to 6 4 3 All substances that can be pipetted by means of a microliter pipette are termed Liquids All others are Non Liquids and have to be processed with the Non Liquid samples procedure 6 4 1 Non Liquids and other sample material Weigh 50 mg 5 mg sample material into a 1 5 or 2 0 ml reaction tube e g by using scalpels or tooth pickers Be aware of cross contaminations gt Proceed with lysis step 6 5 6 4 2 Liquids Vortex the samples thoroughly Use an amount of 300 ul for all liquid samples gt Proceed with lysis step 6 5 6 4 3 Samples for Bacterial Analyse in liquids milk products Use an amount of 1000 ul and centrifuge it for 5 minutes at 5 000 x g Discard the supernatant and use the pellet for DNA extraction gt Proceed with lysis step 5 6 Note Please avoid pipetting milk clots from milk samples into the reaction tube BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 15 of 19 PureMagic BIORON 6 5 Sample Lysis 1 Prepare a mix of Lysis Buffer and Proteinase K For eac
17. tex mixer e Thermomixer and optional Rotator e Magnetic stand e g BIORON Diagnostics GmbH Product No VBE9291 e Analytical Lab Scale e Tabletop centrifuge capable of 16 000 x g e Scalpel or disposables like tooth pickers for the food samples 5 3 Before starting e If in Lysis Buffer a precipitate is formed dissolve it by incubating at 40 50 C till solution is clear e Wash Buffer 10 tests Add 14 ml of 100 Isopropylalcohol to the Wash Buffer before using it for the first time 50 100 tests Add 21 ml of 100 Isopropylalcohol to the Wash Buffer before using it for the first time Note After adding Isopropylalcohol Wash Buffer is stable for a minimum of 1 year at RT e Proteinase K is supplied as a lyophilized powder 10 tests Add 550 ul Dilution Buffer to the lyophilized powder and store the reconstituted Proteinase K at 18 22 C 50 100 tests Add 2 75 ml Dilution Buffer to the lyophilized powder and store the reconstituted Proteinase K at 18 22 C Note After reconstitution Proteinase K has a concentration of 20 mg ml and should be stored at 18 22 C e Pre Heat a thermomixer for incubation at 55 C see 5 6 and later at 65 C see 5 8 e Label all tubes accurately to the laboratory standard BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 8 of 19 PureMagic BIORON 5 4 Bead P
18. to the Increase the time of binding on rotator docile bande Make sure Lysis Buffer does not contain Low DNA yield g precipitates Not enough magnetic beads Make sure beads are sufficiently suspended were used before pipetting pole USO LYSIS ue Incubate the buffer by 40 C 50 C shows precipitates meompicle Resuspend the magnetic beads by resuspension of vortexing before use magnetic beads Magnblit Daada wars Don t freeze or centrifuge the magnetic aggregated they were particles frozen or centrifuged Inefficient cell lysis due to inefficient mixing of Make sure the sample is thoroughly Lysis Buffer and mixed with Lysis Buffer sample No DNA Wash Buffer was not Prepare Wash Buffer by adding eluted prepared correctly Isopropanol according to instruction Repeat lysis step Eluate is not Sample is not completely Centrifuge the solution to separate the DNA clear purified from the impurities and removing the DNA containing supernatant to a fresh tube BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 18 of 19 PureMagic BIORON Problem Cause Corrective action Reduce sample volume weight to 50 DNA is contaminated with Or inhibiting substances Perform PCR with a 1 10 dilution of the eluate Problem with d i with TE or water pai be a Not enough DNA in the Quantify the extracted DNA and use suffic
19. ution Proteinase K should be stored at 18 22 C Magnetic beads should be stored at 2 8 C for long term use Do not freeze During shipment or storage in cool ambient conditions in the Lysis Buffer precipitates may be formed Dissolve such precipitates by incubating at 40 50 C and gently shaking For the shipment temperatures up to 30 C are allowed for a maximum of 7 days 06012015IH BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net Page 6 of 19 PureMagic BIORON 4 Description of the Method Principle DNA is extracted from homogenized food samples by lysis buffers containing chaotropic salts denaturing agents and detergents Subsequently nucleic acid is bound to magnetic particles using appropriate binding buffer Magnetic particles are coated with silica which presents the binding area for DNA and aids as solid support to separate the beads from the solution by a strong magnet The supernatant can be removed and the DNA which stays on the solid bead can further purified by washing Nucleic acids are released in Elution Buffer 5 Extraction Protocol 5 1 General Remarks and Precautions d In the following the individual working steps are described in detail To obtain optimal results it is recommended to read the instruction completely before starting the examination Under laboratory standards safety pre
20. x to each well Transfer the supernatant lysate into the according well Optional using a repeater or a multichannel pipette Washing 1 2 Add 500 pl and 1000 ul Wash Buffer Elution Add 75 ul Elution Buffer BIORON GmbH Rheinhorststr 18 67071 Ludwigshafen Germany Phone 49 621 5720 915 Fax 49 621 5720 916 info bioron net www bioron net 06012015IH Page 16 of 19 BIORON PureMagic Plate Piate Type Content Volume sample Name Tip Plate KingFisher Flex 96 KF Plate Lysate 350 ul 70 Isopropylalcohol Binding Microtiter DW96 Plate lsopropylalcohol 75 ul prepared magnetic beads beads Washing 1 Microtiter DW96 Plate Wash Buffer 500 ul Washing 2 Microtiter DW96 Plate Wash Buffer 1000 ul Elution KingFisher Flex 96 KF Plate Elution Buffer 75 ul Follow instructions on the instruments display and load the prefilled plates in the right position When all plates are loaded start the program Note Ask us for the program with the instrument settings For a seamless processing of samples remember to See KingFisher Flex Instrument User Manual for detailed instructions Combine the Tip Comb with a Deep Well 96 Plate Load the plates into the KingFisher Flex instrument according to the protocol request placing each plate in the same orientation Confirm each action by pressing Start After sample processing remove plates as instructed by the instrument s display
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