120328 Manual Art D-2015
Contents
1. 37 38 42 43 S22 24 26 36 37 39 46 Reagent 8 Poly A RNA no hazardous ingredients Reagent 9 Poly A RNA Buffer Guanidine thiocyanate CAS No 593 84 0 EC No 209 812 1 Xn R20 21 22 32 52 53 S13 61 V120328 Page 6 11 gt Perkin For the Better Equipment and other material to be provided by the user chemagic Prepito RNAse free water disposable gloves pipette and pipette tips with aerosol barrier ensure that all used material is RNase free Purification Protocol using the chemagic Prepito The protocol is suitable for up to 12 samples in parallel see protocol steps below Detailed instructions for the use of the chemagic Prepito can be found in the corresponding user manual Before you start e Check all kit components for integrity In case of damages contact your supplier e Connect the tubes according to their numbering to the respective counterparts at the chemagic 8 Pack Remove the lids from the individual buffer bottles in the chemagic 8 Pack and pierce the septum with the spike at the end of the tube Place the chemagic 8 Pack upside down on the reagent holder e Dissolve the lyophilized Protease in RNAse free water see instruction on the tube and Poly A RNA in 440 uL Poly A RNA Buffer per tube Positioning of the Deep Well Plate and the chemagic Tip amp Tube Rack The following scheme shows the orientation of the 96 Deep Well Plate and the chemagic Tip amp Tube Rack For detailed information se2. Prepito Viral DNA RNA D200 Kit art No D 2015 CE gt PerkinElmer gt Perkin For the Better Symbols y So Kit contains reagents for 180 preparations Ci Refer to information given in the handbook V120328 Expiry date LOT Lot number IVD in vitro diagnostic medical device X Temperature limitations REF D 2015 ul PerkinElmer chemagen GmbH Arnold Sommerfeld Ring 2 D 52499 Baesweiler Tel 49 2401 805500 Intended Use With the Prepito Viral DNA RNA D200 Kit viral nucleic acids can be isolated from plasma or serum for subsequent in vitro diagnostic purposes The Kit has to be used with the chemagic Prepito The product is intended for professional users such as technicians and physicians trained in molecular biology techniques To minimize irregularities in diagnostic results the product should always be used with an internal control as well as positive and negative controls throughout the process of sample preparation sample amplification and detection according to the downstream assay used Any diagnostic results generated using the sample preparation procedure in conjunction with any downstream diagnostic NAT assay should be interpreted with regard to other clinical or laboratory findings V120328 Page 2 11 Content SYMDO S c ri iaia 2 hended USE ili lia lie i aa 2 Funetionaliprinciple iisade eil laniaat 4 Quality Control aaa iaia 4 Product Ilmitations cc ale eni ae ML e
3. ange Protocol Press Serum Plasma in the Select Protocol Group window Select the Prepito Viral DNA RNA200 Kit Kit protocol by pressing Viral DNA RNA 200 and confirm by pressing OK Confirm the protocol selection in the Select Protocol Group window by pressing OK Enter the 4 digit access code 2365 for authorization and confirm by pressing Enter Press Start Process Read the protocol information in the appearing information screen and confirm by pressing Continue Select the sample positions and confirm by pressing OK Enter the kit barcode with the barcode scanner and confirm by pressing OK For the registration of the samples and elution tubes press Yes and follow the instructions on the touch screen panel to enter the according barcodes Prepare the chemagic Tip amp Tube Rack with the required material Place one 0 75 mL reaction tube filled with 50 100 uL Elution Buffer position 1 one 0 75 mL reaction tube filled with 150 uL Magnetic Beads position 2 and one Disposable Tip position 3 for each sample into the positions according to the sample positions Shake the vessel with the Magnetic Beads vigorously until all Magnetic Beads are completely resuspended An incomplete resuspension of the Magnetic Beads can result in a decreased yield of extracted nucleic acids Add 4 uL Poly A RNA solution and 10 uL Protease solution into the sample position of the Deep Well DWP defined as sample wells Pos lys
4. ate mix see section above Positioning Procedure Add 200 uL sample material and 200 uL Lysis Buffer into the well filled with Poly A RNA and Protease solutions An information screen indicates the previously selected sample positions Ensure that the sample positions in the DWP correspond to the selected positions Place the DWP on its default position on the tracking system and press Continue Place the chemagic Tip amp Tube Rack on its default position on the tracking system Check for accurate fit of the DWP and the chemagic Tip amp Tube Rack and lock both by closing the safety latch Close the front door and immediately start the automated isolation process by pressing Start Page 9 11 gt Perkin For the Better General remarks It is strongly recommended to use the extracted nucleic acids immediately for amplification If nucleic acid extracts cannot be used for amplification directly after preparation the nucleic acid extracts can be kept at 20 or preferably at 70 for up to one month or one year respectively The Elution Buffer included in this kit is 10 mM Tris HCI pH 8 0 V120328 Page 10 11 Troubleshooting gt Perkin For the Better Problem Possible Cause Recommendation Solution Red eluates low detection sensitivity Traces of erythrocytes in the plasma or serum samples Avoid to carry over erythrocytes during the preparation of plasma or serum Low detection s
5. e protocols steps amp oe Qa Vv xe Co O 200 uL sample material Protease and Poly A RNA 2 J O O O oO Pos 4 second row for Disposable Tips not used in this protocol ag 0O O O DO Pos 3 Disposable Tips 2 amp JOOO O Pos 2 0 75 mL reaction tubes with 150 uL Magnetic Beads Oo POO0O O Pos 1 0 75 mL reaction tubes with 50 100 uL Elution Buffer V120328 Page 7 11 gt Perkin For the Better Protocol Steps chemagic Prepito serial numbers 1 99 1 2 3 4 5 6 10 11 12 13 14 15 V120328 Switch on the chemagic Prepito and wait for the self test to finish Press change protocol Select the Prepito Viral DNA RNA D200 Kit protocol by pressing Viral DNA RNA 200 Enter the access code 2365 for authorization and confirm by pressing enter Confirm the selection of the correct protocol by pressing enter Read the protocol information in the appearing information screen Confirm by pressing continue Select the sample positions and confirm by pressing continue Enter the kit barcode with the barcode scanner and confirm by pressing ok For the registration of the samples and storage tubes press yes and follow the instructions on the touch screen panel to enter the according barcodes Prepare the chemagic Tip amp Tube Rack with the required materials Place one 0 75 mL reaction tube filled with 50 100 uL Elu
6. ee ee 4 Stability and storage sia oo RIE Le Lee eee 4 Protocol GUATO ainsuse an iero 4 Contents of the Kit unit corresp to 180 preparations from 200 uL serum or plasma 5 ALCL attic lia ia ei 6 Equipment and other material to be provided by the USE 7 Purification Protocol using the chemagic Prepito nenian annia ania ananin aaaaa na 7 Positioning of the Deep Well Plate and the chemagic Tip amp Tube Rack i 7 Protocol Steps chemagic Prepito serial numbers 1 99 8 Protocol Steps chemagic Prepito serial numbers 100 and later i 9 GON eral remark Sirosis aa Rin 10 Troubleshooting cc criari R RAEE aaa 11 V120328 Page 3 11 gt Perkin For the Better gt Perkin For the Better Functional principle The chemagic Prepito Viral DNA RNA D200 Kit is based on chemagen s proprietary magnetic bead technology platform Viruses in the sample material are lysed during the isolation process The released nucleic acids bind to small magnetisable particles which are then magnetically separated from the sample material During subsequent steps contaminations are removed and the purified nucleic acids are transferred into an elution medium The automated sample processing by the chemagic Prepito excludes cross contamination and ensures a safe handling of infectious sample material Quality control Each lot is tested for its defined specif
7. ensitivity for positive controls and or target nucleic acid Incorrect amount of Magnetic Beads added Resuspend the Magnetic Beads well before adding to the lysate Insufficient lysis Add the correct volume of lysis buffer Buffers in the chemagic 8 Pack are not connected to the machine Connect the buffers in the chemagic 8 Pack to the machine The chemagic 8 Pack is not positioned in the right manner on the reagent holder Place the chemagic 8 Pack in the correct position on the reagent holder Tubes contain air after connecting the chemagic 8 Pack to the machine Fill the tubes completely using the manual priming function Buffers in the chemagic 8 Pack are empty Change the chemagic 8 Pack Don t use the chemagic 8 Pack for more than the indicated preparations Buffers of the chemagic 8 Pack are not connected in the right manner to the chemagic Prepito Check correct the connections between the chemagic Prepito and the chemagic 8 Pack Irregular dispensing of the buffers Check the calibration of the pumps Contaminated or inactive Protease Visible microbial growth in Protease solution Use sterile water for resuspension of the Protease Incorrect storage of the Protease solutions Store Protease solution at 4 do not use the solutions longer than 6 weeks Store aliquots at 20 Avoid thawing freezing cycles Malfunction of the instrument e g mechanica
8. ffer 2 130 mL Tris HCl buffer Sodium perchlorate 25 28 Ethanol 45 60 Wash Buffer 3 110 mL Tris HCl Puffer Sodium perchlorate 15 18 Ethanol 20 25 Wash Buffer 4 110mL Ethanol 70 80 Elution Buffer 5 30 mL 10 mM Tris HCl buffer pH 8 0 Protease 2 0 mL Poly A RNA 2 x 350 ug Poly A RNA Buffer 2 x 440 uL Disposable Tips 180 2 mL Deep Well Plates 15 0 75 mL Reaction Tubes 360 0 75 mL Caps 180 included in the chemagic 8 Pack Page 5 11 For the Better gt PerkinElmer Safety To avoid injuries while working with the kit components always wear safety glasses disposable gloves and protective clothing For detailed information please refer to the according material safety data sheet MSDS Reagent 1 Magnetic Beads no hazardous ingredients Reagent 2 Lysis Buffer 1 Guanidine thiocyanate CAS No 593 84 0 EC No 209 812 1 Xn R20 21 22 32 52 53 S13 61 Reagent 3 Binding Buffer 2 Sodium perchlorate CAS No 7601 89 0 EC No 231 511 9 Xn R9 22 S13 22 27 Ethanol CAS No 64 17 5 EC No 200 578 6 F R11 Reagent 4 Wash Buffer 3 Ethanol CAS No 64 17 5 EC No 200 578 6 F R11 S7 16 Sodium perchlorate CAS No 7601 89 0 EC No 231 511 9 Xn R9 22 S13 22 27 Reagent 5 Wash Buffer 4 Ethanol CAS No 64 17 5 EC No 200 578 6 F R11 S7 16 Reagent 6 Elution Buffer 5 no hazardous ingredients Reagent 7 Protease Protease CAS No 9036 06 0 EC No 232 909 5 Xn R36
9. ications according to chemagen s Quality Management System Procedures that are not in accordance with this manual could cause inadequate results Product limitations The Kit is designed for the use with humane plasma or serum The Kit is not intended for the use with tissue or blood sample material The isolation efficiency with other types of sample material has not been determined Stability and storage Expiry dates are stated on the box and the single components of the kit Do not use any components of the kit beyond the expiry date All kit components can be stored at room temperature Lysis Buffer 1 plasma and Poly A RNA Buffer have to be stored in the dark Lysis Buffer 1 may form a precipitate upon storage If necessary warm to approximately 55 to redissolve Precipitates in the Poly A RNA buffer can be redissolved at room temperature After reconstitution Protease solution and Poly A RNA solution have to be stored at 4 The solutio ns can be used for 6 weeks For long term storage we recommend aliquoting the Proteinase K solution and the Poly A RNA solution and storing at 20 Protocol duration The length of the purification protocol is 70 min V120328 Page 4 11 For the Better gt Perkin Contents of the Kit unit corresp to 180 preparations from 200 uL serum or plasma i 2 10 11 12 13 V120328 Magnetic Beads 30 mL Lysis Buffer 1 45 mL Guanidine thiocyanate 43 50 Binding Bu
10. l electrical or electronical problems Contact chemagen or your local supplier detailed information is given in the manual of the chemagic Prepito V120328 Page 11 11
11. tion Buffer position 1 one 0 75 mL reaction tube filled with 150 uL of Magnetic Beads position 2 and one disposable tip position 3 for each sample into positions according to the sample positions Shake the Magnetic Bead solution vigorously until all Magnetic Beads are completely suspended An incomplete resuspension of the Magnetic Bead solution could cause a decreased yield of extracted nucleic acids Add 4 uL Poly A RNA solution and 10 uL Protease into the sample position of the Deep Well Plate DWP riplate SW Add 200 uL sample material and 200 uL Lysis Buffer 1 into the well prefilled with Poly A RNA and Protease solutions An information screen indicates the previously selected sample positions Ensure that the sample positions in the DWP correspond to the selected positions Place the DWP on its default position on the tracking system and press continue Place the chemagic Tip amp Tube Rack on its default position on the tracking system Check the accurate fit of the DWP and chemagic Tip amp Tube Rack and lock both by closing the safety latch Close the front door and immediately start the automated isolation process by pressing start Page 8 11 gt Perkin For the Better Protocol Steps chemagic Prepito serial numbers 100 and later 1 A DN DS ol 10 11 12 13 14 15 16 17 V120328 Switch on the chemagic Prepito and wait until the self test is finished Press Ch
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