User manual / Technical Information
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1. Avoid repeated freeze amp thaw cycles for the tissue to be extracted All materials and reagents used for DNA or RNA isolation should be free of DNases or RNases 4 General Comments on Handling of RNA RNA is far less stable than DNA It is very sensitive to degradation by endogenous RNases in the biological material and exogenous RNases which are permanently present everywhere in the lab In order to achieve satisfactory qualitative and quantitative results in RNA preparations contaminations with exogenous RNases have to be reduced to a minimum in accordance with the following recommendations Clear the bench top first using RNase AWAY A998 1 or Roti Nucleic Acid free HP69 1 Always wear latex or vinyl gloves while handling reagents and RNA samples in order to prevent RNase contaminations from surface of the skin or from dusty laboratory equipment Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI 6 Use only sterile disposable polypropylene tubes throughout the procedure these tubes are generally RNase free Autoclaving will not inactivate RNases Non disposable plastic ware should be treated before use to ensure that it is RNase free Plastic ware should be thoroughly rinsed with RNase AWAY A998 1 or Roti Nucleic Acid free HP69 1 followed by RNase free water All glassware should be treated before use in order to ensure that it is RNase free Glassware should be cle2. 5 Elution Place the Spin Column into a clean 1 5 ml elution tube Add 60 l pre heated DNase free or RNase free water incl in the kit to the centre of the membrane Incubate for 2 mins at room temperature Centrifuge at 8 000 g or 10 000 rpm for 1 min RT to elute DNA RNA We recommend using a shaking platform or thermomixer for continuous agitation of the sample Alternatively vortex the sample 3 4 times during incubation Agitation is vital for a decent recovery rate If the solution has not completely passed through the spin column centrifuge again at higher speed or prolong the centrifugation time The type of water to be used depends on whether viral DNA or viral RNA shall be isolated Elution with lower volumes of water e g 20 30 l increases the final concentration of DNA RNA Higher volumes 80 100 l result in lower concentration of DNA RNA although the overall amount of DNA RNA eluted may be slightly higher Repetition of the elution step may increase the yield of DNA RNA overall but may reduce the concentration after pooling of the fractions Minimum elution amount is 20 l Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 9 5 2 Isolation of viral DNA RNA from 400 l cell free fluids Step RT room temperature done 1 Virion Lysis Pipet 400 l Mix of Lysis Buffer LSV Carrier CS into a 2 0 ml reaction tube Add 400 l of the sample and 20 l Proteinase K solution Mix vigorously
3. USER MANUAL Roti Prep Viral DNA RNA MINI For isolation of viral DNA and RNA from various sources Ord No 8547 Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI For isolation of viral DNA and RNA from a variety of sample materials Introduction and product description Preparation by well known mini spin column system Fast easy and efficient For isolation of viral ssDNA dsDNA and ssRNA Preparation in approx 25 minutes Elution in 60 l Roti Prep Viral RNA DNA MINI has been developed for the isolation of viral DNA and RNA from various kinds of source material The isolation is carried out via the proven and reliable spin column technique which is very easy to handle and needs only few steps The kit enables the simultaneous extraction of viral ssDNA dsDNA and ssRNA The purification happens by using a carrier RNA which engenders a higher binding of the nucleic acid resp its complex with the carrier RNA on the spin filter and thus a higher recovery rate Furthermore in order to achieve a maximum of nucleic acid purity for each source material 5 specifically optimised protocols are provided Thus within approx 25 mins high pure viral nucleic acid is eluted free from inhibitors for further downstream applications like RT PCR PCR packaging etc The Kit has been successfully tested for e g HBV CMV EBV HSV FMDV Suitable source material T
4. Your notes Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI 16 Ordering information for detailed kit content see Table under 1 To place your order please contact us Phone Germany 0800 56 99 000 Phone international 49 0 721 5606 0 Fax Germany and international 49 0 721 5606 149 E Mail Germany bestellungen carlroth de E Mail international info carlroth com Product number Product Content Amount 8547 1 Roti Prep Viral DNA RNA MINI 1 Kit 50 Preps Carl Roth GmbH Co KG Schoemperlenstra e 3 5 76185 Karlsruhe Germany Phone 49 0 721 5606 0 Fax 49 0 721 5606 149 Email info carlroth com
5. 10 000 g or 12 000 rpm for 1 min RT and discard the flow through 3 Column Washing Place the Spin Column into a new collection tube Add 500 l of Washing Buffer WSA to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Again centrifuge columns at 10 000 g or 12 000 rpm for 5 mins RT in order to remove residual ethanol Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 11 Sequel Isolation of viral DNA RNA from tissue samples or biopsies Step RT room temperature done 5 Elution Place the Spin Column into a clean 1 5 ml elution tube Add 60 l pre heated DNase free or RNase free water incl in the kit to the centre of the membrane Incubate for 2 mins at room temperature Centrifuge at 8 000 g or 10 000 rpm for 1 min RT to elute DNA RNA We recommend using a shaking platform or thermomixer for continuous agitation of the sample Alternatively vortex the sample 3 4 times during incubation Agitation is vital for a decent recovery rate If the solution has not completely passed through the spin column centrifuge again at higher speed or prolong the centrifugation t
6. addition of cold distilled sterile RNase free H2O as follows 8547 1 50 Preps 1 5 ml Mix thoroughly Dissolve Carrier CS by addition of cold distilled sterile RNase free H2O as follows 8547 1 50 Preps 1 25 ml Mix thoroughly by pipetting Dissolved Carrier CS should be stored in aliquots at 20 C Do not repeatedly freeze amp thaw 3times in maximum Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 7 Add the appropriate amount of Carrier CS to an aliquot of Lysis Buffer LSV Lysis Buffer Carrier Mix for isolation of nucleic acid from 200 l cell free fluid Component 1 sample 4 samples 10 samples Lysis Buffer LSV 240 l 960 l 2 4 ml Carrier CS 12 l 48 l 120 l Final vol 252 l 1008 l 2 52 ml Lysis Buffer Carrier Mix for isolation of nucleic acid from 400 l cell free fluid Component 1 sample 4 samples 10 samples Lysis Buffer LSV 480 l 1920 l 4 8 ml Carrier CS 12 l 48 l 120 l Final vol 492 l 1968 l 4 92 ml Mixture of Lysis Buffer LSV and Carrier CS is stable for one day at 4 C Heat one thermal mixer or water bath to 50 C Preheat one tube of RNase free water to 70 C Please note Centrifugation steps should be carried out at room temperature 5 Workflow The workflow has been provided on a separate page in order to use a copy for in process protocol or filing in your personal lab files Carl Roth GmbH Co KG Roti Prep V
7. by pulsed vortexing for 10 sec Incubate at 70 C under constant agitation for 10 mins Centrifuge the lysed sample at 10 000 g or 12 000 rpm for 30 sec to remove condensate from the lid of the tube Add 800 l Binding Buffer BSN to the lysed sample Mix by vortexing or pipetting until a homogenous solution is achieved 2 Column Loading Place each blue Spin Column into a 2 ml collection tube Apply 650 l of the mix of lysed sample Binding Buffer BR to the spin column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Apply residual mix of lysed sample Binding Buffer BR to the spin column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through 3 Column Washing Place the Spin Column into a new collection tube Add 500 l of Washing Buffer WSA to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Again centrifuge columns at 10 000 g or 12 000 rpm for 5 mins RT in order to remove residual ethanol 5 Elution Place the Spin Column into a clean 1 5 ml elution tube Add 60 l pre heated DNase free or RNase free water inc
8. 00 l Binding Buffer BSN to the lysed sample Mix by vortexing or pipetting until a homogenous solution is achieved 2 Column Loading Place each blue Spin Column into a 2 ml collection tube Apply the mix of lysed sample Binding Buffer BSN to the spin column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through 3 Column Washing Place the Spin Column into a new collection tube Add 500 l of Washing Buffer WSA to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Again centrifuge columns at 10 000 g or 12 000 rpm for 5 mins RT in order to remove residual ethanol 5 Elution Place the Spin Column into a clean 1 5 ml elution tube Add 60 l pre heated DNase free or RNase free water incl in the kit to the centre of the membrane Incubate for 2 mins at room temperature Centrifuge at 8 000 g or 10 000 rpm for 1 min RT to elute DNA RNA We recommend using a shaking platform or thermomixer for continuous agitation of the sample Alternatively vortex the sample 3 4 times during incubation Agitation is vital for a decent recovery rate If the solution has
9. O DNase free e g HN57 1 sterilize in an autoclave Distilled sterilized H2O RNase free for preparation of 70 ethanol e g T143 1 1x PBS DNase free or RNase free e g Roti Stock 10x PBS 1059 1 0 9 NaCl solution RNase free e g HN00 NaCl powdered CELLPURE with low endotoxin content DEPC e g K028 1 3 Application Roti Prep Viral DNA RNA MINI is designed for isolation of high purity viral DNA and or RNA from various source material Spin column based preparation allows elution in a small volume of low salt buffer eliminating time consuming phenol chloroform extraction or alcohol precipitation The columns may be used either in micro centrifuges or on vacuum manifolds The detection limit for certain viruses depends on the individual procedures for example in house PCR or commercial detection assays used The Roti Prep Viral DNA RNA MINI kit contains a Carrier Mix Carrier CS with Carrier RNA as well as an internal control IC DNA and RNA This IC DNA RNA may be used for checking the success of DNA RNA isolation by real time PCR Use of the Carrier CS is optional Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 5 Please note that in case of using the Carrier CS the eluate contains isolates viral nucleic acids as well as IC DNA RNA and Carrier RNA In this case quantitation of the isolated nucleic acids by photometric or fluorometric methods is not possible We recommend to qu
10. aned with RNase AWAY A998 1 or Roti Nucleic Acid free HP69 1 thoroughly rinsed with RNase free water and oven baked at 240 C for four or more hours before use Oven baking in activates RNases and ensures that no other nucleic acids such as plasmid DNA are present on the surface of the glassware Reduce preparation time as much as possible Change gloves frequently and keep tubes closed Keep isolated RNA on ice Electrophoresis tanks should be cleaned with RNase AWAY A998 1 or Roti Nucleic Acid free HP69 1 followed by RNase free water rinsed with ethanol and finally allowed to dry All buffers have to be prepared with DEPC treated RNase free double distilled water Avoid handling bacterial cultures cell cultures or other biological sources of RNases in the same lab where the RNA purification will be performed Do not use equipment glassware and plastic ware employed for other applications which might introduce RNase contaminations in the RNA isolation Before start be sure to Add Ethanol 96 99 8 not incl in this kit to the Washing Buffer WSA as follows 8547 1 50 Preps 15 ml 30 ml final vol Mix thoroughly and keep the bottle always firmly closed Add Ethanol 96 99 8 not incl in this kit to the Washing Solution WSL as follows 8547 1 50 Preps 64 ml 80 ml final vol Mix thoroughly and keep the bottle always firmly closed Dissolve Proteinase K by
11. antify extracted DNA and RNA by other methods like specific quantitative PCR or real time PCR Additionally Carrier RNA may inhibit PCR reactions Thus the amount of Carrier RNA added has to be carefully optimized depending on the individual PCR system used for down stream analyses As in all isolation procedures lysis is a crucial step and has to be performed very carefully Complete lysis of the sample is vital for a good recovery rate of nucleic acid Generally virus lysis is easily achieved under the conditions given in the protocols However if the recovery rate is not as good as expected lysis conditions may have to optimised nevertheless Please focus on the fact that RNA is highly temperature sensitive and has to be kept cool as soon as possible It is important therefore to prolong all steps as long as necessary but also as short as possible The lysed samples may be stored prior to adding the Binding Buffer Store samples of RNA virions at 80 C and samples of DNA virions at 20 Repetition of elution final step may the recovery rate overall However even so often concentration of the recovered RNA is reduced in parallel since the whole amount of water used for elution is higher Store the extracted DNA at 20 C extracted RNA at 80 C Avoid repeated freeze amp thaw cycles by all means For centrifugation we recommend a standard microcentrifuge Centrifugation steps should be carried out at room temperature
12. ime The type of water to be used depends on whether viral DNA or viral RNA shall be isolated Elution with lower volumes of water e g 20 30 l increases the final concentration of DNA RNA Higher volumes 80 100 l result in lower concentration of DNA RNA although the overall amount of DNA RNA eluted may be slightly higher Repetition of the elution step may increase the yield of DNA RNA overall but may reduce the concentration after pooling of the fractions Minimum elution amount is 20 l Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI 12 5 4 Isolation of viral DNA RNA from swabs Step RT room temperature done 1 Virion Lysis Place the swab into a 1 5 ml reaction tube and add 0 9 NaCl solution DNase or RNase free so that the swab head is completely covered Incubate at RT for 15 mins Shake the swab vigorously Then remove it from the tube by pressing the tip of the swab to the inner tube wall in order to quantitatively squeezing the liquid from the cotton wool into the tube This solution is the sample used in the following steps Pipet 200 l Mix of Lysis Buffer LSV Carrier CS into a 1 5 ml reaction tube Add 200 l of the sample and 20 l Proteinase K solution Mix vigorously by pulsed vortexing for 10 sec Incubate at 70 C under constant agitation for 10 mins Centrifuge the lysed sample at 10 000 g or 12 000 rpm for 30 sec to remove condensate from the lid of the tube Add 4
13. iral DNA RNA MINI 8 5 1 Isolation of viral DNA RNA from 200 l cell free fluids Step RT room temperature done 1 Virion Lysis Pipet 200 l Mix of Lysis Buffer LSV Carrier CS into a 2 0 ml reaction tube Add 200 l of the sample and 20 l Proteinase K solution Mix vigorously by pulsed vortexing for 10 sec Incubate at 70 C under constant agitation for 10 mins Centrifuge the lysed sample at 10 000 g or 12 000 rpm for 30 sec to remove condensate from the lid of the tube Add 400 l Binding Buffer BSN to the lysed sample Mix by vortexing or pipetting until a homogenous solution is achieved 2 Column Loading Place each blue Spin Column into a 2 ml collection tube Apply the mix of lysed sample Binding Buffer BSN to the spin column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through 3 Column Washing Place the Spin Column into a new collection tube Add 500 l of Washing Buffer WSA to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Add 650 l of Washing Solution WSL to the Spin Column Centrifuge at 10 000 g or 12 000 rpm for 1 min RT and discard the flow through Again centrifuge columns at 10 000 g or 12 000 rpm for 5 mins RT in order to remove residual ethanol
14. issue up to 20 mg Biopsies up to 20 mg Supernatant medium from cell culture up to 400 l Serum plasma or liquor up to 400 l Cell free body fluids up to 400 l Swabs For research use only Information in this document is subject to change without notice Carl Roth GmbH Co KG assumes no responsibility for any errors that may appear in this document Carl Roth GmbH Co KG disclaims all warranties with respect to this document expressed or implied including but not limited to those of merchantability or fitness for a particular purpose In no event shall Carl Roth GmbH Co KG be liable whether in contract tort warranty or under any statute or on any other basis for special incidental indirect punitive multiple or consequential damages in connection with or arising from this document including but not limited to the use thereof Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 3 Cautions Binding Buffer BSN Danger H225 H318 H336 H411 P210 P280 P303 P361 P353 P305 P351 P338 P312 Lysis Buffer LSV Danger H318 P280 P305 P351 P338 P310 Proteinase K Danger H315 H319 H334 H317 H335 P280 P284 P333 P313 P337 P313 P342 P311 Washing Buffer WSA Warning H302 H332 H412 EUH032 P301 P312 P304 P340 In case of forming of precipitates please dissolve by careful warming MSDS the appropriate MSDS can be downloaded from our website www carlroth com All due care and attention
15. l in the kit to the centre of the membrane Incubate for 2 mins at room temperature Centrifuge at 8 000 g or 10 000 rpm for 1 min RT to elute DNA RNA We recommend using a shaking platform or thermomixer for continuous agitation of the sample Alternatively vortex the sample 3 4 times during incubation Agitation is vital for a decent recovery rate If the solution has not completely passed through the spin column centrifuge again at higher speed or prolong the centrifugation time The type of water to be used depends on whether viral DNA or viral RNA shall be isolated Elution with lower volumes of water e g 20 30 l increases the final concentration of DNA RNA Higher volumes 80 100 l result in lower concentration of DNA RNA although the overall amount of DNA RNA eluted may be slightly higher Repetition of the elution step may increase the yield of DNA RNA overall but may reduce the concentration after pooling of the fractions Minimum elution amount is 20 l Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI 10 5 3 Isolation of viral DNA RNA from tissue samples or biopsies max 20 mg Step RT room temperature done 1a Tissue Lysis with a Homogeniser Place max 20 mg of the tissue sample fresh or frozen material into a suitable vessel for homogenisation Add 200 500 l DNase free or RNase free buffer water or PBS and homogenise the sample to fine powder according to instructions acco
16. mpanying the instrument used Transfer the homogenised tissue buffer mix to a 1 5 ml reaction tube Centrifuge the mixture at 15 000 g or maximum speed for 2 mins in order to remove unhomogenised material 1b Tissue Lysis with Mortar and Pestle in Liquid Nitrogen Place max 20 mg of the tissue sample fresh or frozen material into a suitable vessel for grinding e g a mortar When using fresh material add liquid nitrogen for freezing Grind the sample with a pestle to fine powder Add 200 500 l DNase free or RNase free buffer water or PBS and mix thoroughly Transfer the homogenised sample to a 1 5 ml reaction tube Centrifuge the mixture at 15 000 g or maximum speed for 2 mins in order to remove unhomogenised material 2 Virion Lysis Pipet 200 l Mix of Lysis Buffer LSV Carrier CS into a 1 5 ml reaction tube Add 200 l of the supernatant of the centrifuged sample Also add 20 l Proteinase K solution Mix vigorously by pulsed vortexing for 10 sec Incubate at 70 C under constant agitation for 10 mins Centrifuge the lysed sample at 10 000 g or 12 000 rpm for 30 sec to remove condensate from the lid of the tube Add 400 l Binding Buffer BSN to the lysed sample Mix by vortexing or pipetting until a homogenous solution is achieved 3 Column Loading Place each blue Spin Column into a 2 ml collection tube Apply the mix of lysed sample Binding Buffer BSN to the spin column Centrifuge at
17. not completely passed through the spin column centrifuge again at higher speed or prolong the centrifugation time The type of water to be used depends on whether viral DNA or viral RNA shall be isolated Elution with lower volumes of water e g 20 30 l increases the final concentration of DNA RNA Higher volumes 80 100 Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 13 l result in lower concentration of DNA RNA although the overall amount of DNA RNA eluted may be slightly higher Repetition of the elution step may increase the yield of DNA RNA overall but may reduce the concentration after pooling of the fractions Minimum elution amount is 20 l Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI 14 6 Trouble Shooting Problem probable cause Comments and suggestions Clogged spin filter Insufficient lysis and or too much starting material Increase lysis time or centrifugation speed After lysis centrifuge the lysate to pellet unlysed material Continue with the supernatant Check whether the protocol for virus lysis has been applied Reduce amount of starting material Overload of filters reduces yield Low recovery amount of isolated nucleic acid Insufficient lysis See above Incomplete elution Prolong incubation time with DNase free or RNase free water to 5 minutes or repeat elution step once again Use higher volume of DNase free RNase free water for elution Insufficient mi
18. should be exercised in handling the materials and reagents contained in the kit Always wear gloves while handling these reagents and avoid any skin contact In case of contact flush eyes or skin with a large amount of water immediately Research samples gained from patient s material like serum liquor etc sample must always be considered as potentially infectious Samples from risk patients must always be labelled and handled under consequent safety conditions Please observe the federal state and local safety and environmental regulations Literature Citation When describing a procedure for publication using this product please refer to it as Carl Roth s Roti Prep Viral DNA RNA MINI Kit 1 Materials provided with the kit and storage conditions Amount Component Storage 30 mg Proteinase K 4 C 20 C 25 ml Lysis Buffer LSV gt 20 C 60 ml Binding Buffer BSN RT 0 5 mg Carrier CS Base 20 C 3x2 ml RNase free water RT 15 ml Washing Buffer WSA Base RT 16 ml Washing Solution WSL Base RT 50 Mini spin columns blue RT 50 1 5 ml Elution tubes RT 6x50 2 ml Collection tubes RT Carl Roth GmbH Co KG Roti Prep Viral DNA RNA MINI 4 Lyophilized Proteinase K should be stored at 4 C Prior to use dissolve Proteinase K in 1 5 ml sterile distilled water as given below Dissolved Proteinase K should be stored in aliquots at 20 C Avoid repeated freeze amp thaw c
19. xing with Binding Buffer BSN Mix sample with Binding Buffer BSN by pipetting or by vortexing prior to transfer of the sample onto the Spin Filter Low concentration of isolated nucleic acid Too much RNase free Water Elute the viral DNA RNA with a lower volume of DNase free RNase free water No Carrier CS added Add Carrier RNA Carrier CS to the sample as described above Degraded RNA RNA source inappropriately handled or stored See above Check condition of the starting material Is it fresh and undamaged RNA is very temperature sensitive Make sure the protocol especially the first steps are performed as long as necessary but as short as possible RNase contamination of solutions Collection Tubes etc Use sterile RNase free filter tips Before every preparation clean up the pipette the devices and the working place Always wear gloves Old material Use fresh material Check and improve storage conditions Incorrect storage of starting material Avoid repeated freezing and thawing of the starting material Viral RNA does not perform well in downstream applications e g RT PCR Ethanol carryover during elution Increase time for removing of ethanol Salt carryover during elution Ensure that Washing Buffer WSA and Washing Solution WSL are at room temperature Check Washing Solutions for salt precipitates Dissolve these by careful warming Roti Prep Viral DNA RNA MINI Carl Roth GmbH Co KG 15
20. ycles for each tube Lysis Buffer should be stored at over 20 C in order to prevent precipitation of reagents Lyophilized Carrier CS should be stored at 20 C after receipt of the Kit Prior to use dissolve Carrier CS in 1 25 ml sterile RNase free water as given below Dissolved Carrier CS should be stored in aliquots at 20 C Avoid repeated freeze amp thaw cycles for each tube Mixture of Lysis Buffer LSV and Carrier CS is stable for one day at 4 C The Roti Prep Viral DNA RNA MINI Kit should be stored dry at room temperature 14 25 C Before every use make sure that all components have room temperature If there are any precipitates within the provided solutions solve these precipitates by careful warming This kit is stable for at least 6 months after receipt when stored as directed Contents of this Kit may not be bought separately The components of each Roti Prep Viral DNA RNA MINI Kit were tested in general by isolation of viral DNA and RNA and subsequent detection of the internal control DNA and RNA by real time PCR The user is responsible however to validate the performance of the Roti Prep Viral DNA RNA MINI Kit for isolation of RNA from his particular samples since the performance characteristics of our kits have not been validated for any specific application 2 Required Material and Equipment not included in this kit Ethanol 96 99 8 e g Ethanol p a e g 9065 1 Distilled sterilized H2
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