NucleoSpin® DNA Trace - MACHEREY
Contents
1. 18 MACHEREY NAGEL 06 2014 Rev 072. Genomic DNA from forensic samples User manual NucleoSpin DNA Trace June 2014 Rev 07 MACHEREY NAGEL www mn net com Genomic DNA from forensic samples Protocol at a glance Rev 07 Funnel NucleoSpin DNA Trace 1 Lyse sample 4 8 mL FLB 50 uL Proteinase K 56 C 1h 2 Clarify sample e 2 5 000 x g 10 min 3 Adjust DNA binding sonditians 3 5 mL ethanol Vortex 4 Bind DNA am il Load sample y 3 000 x g eS 3 min 5 Wash silica am membrane 1 wash 2 5 mLBW li 2 wash 5 mL B5 V 3 wash 5 mL BS 4st and gro en 9 6 Dry silica 3 000 x g membrane 5 10 min 7 Elute DNA 100 uL BE 70 C RT 2 min e 3 000 x g 3 min MACHEREY NAGEL GmbH amp Co KG Neumann Neander Str 6 8 52355 D ren Germany Tel 49 24 21 969 270 Fax 49 24 21 969 199 tech bio mn net com www mn net com Genomic DNA from forensic samples Table of contents 1 Components 4 1 1 Kit contents 4 1 2 Reagents consumables and equipment to be supplied by user 5 1 3 About this user manual 5 2 Product description 5 2 1 The basic principle 2 2 Kit specifications 3 Storage conditions and preparation of working solutions 8 4 Safety instructions 9 5 Protocols 11 5 1 Isolation of genomic DNA from solid samples for example small amounts of cells or tissue forensic samples 11 5 2 Isolation of genomic DNA from human bones 13 6 Appendix 15 6 1 Troubleshooting 15
3. to the sample and vortex vigorously 5 Bind DNA For each sample take one NucleoSpin DNA Trace F Column placed in a Collection Tube 50 mL Apply the sample successively to the column Centrifuge for 3 min at 3 000 x g Discard the flow through and place the column back into the Collection Tube 6 Wash silica membrane Add 3 mL Buffer BW Centrifuge for 3 min at 3 000 x g Discard the flow through and place the column back into the Collection Tube Add 3 mL Buffer B5 to the column and centrifuge for 3 min at 3 000 x g Discard the flow through and place the column back into the Collection Tube Add 3 mL Buffer B5 to the column and centrifuge for 3 min at 3 000 x g Discard the flow through and place the column back into the Collection Tube 7 Drysilica membrane Centrifuge the column for 10 min at 3 000 x g Residual ethanol is removed during this step 8 Elute highly pure DNA Attach the supplied Elution Tube with adaptor to the NucleoSpin DNA Trace F Column and insert assembly into a new 50 mL centrifuge tube not provided Add 60 uL Buffer BE preheated to 70 C Incubate at room temperature for 2 min Centrifuge for 3 min at 3 000 x g to collect the nucleic acid containing fraction Remove the elution tube containing the nucleic acids and keep it for further use 14 MACHEREY NAGEL 06 2014 Rev 07 Genomic DNA from forensic samples 6 Appendix 6 1 Troubleshooting Problem Possible cause and suggestions Incomplete sa
4. 6 2 Ordering information 16 6 3 Product use restriction warranty 16 MACHEREY NAGEL 06 2014 Rev 07 3 Genomic DNA from forensic samples 1 Components 1 1 Kit contents NucleoSpin DNA Trace 4 preps 25 preps REF 740942 4 740942 25 Lysis Buffer FLB 50 mL 250 mL Wash Buffer BW 13 mL 75 mL Wash Buffer B5 Concentrate 12 mL 100 mL Elution Buffer BE 13 mL 13 mL NucleoSpin DNA Trace F Columns 4 25 plus Collection Tubes Proteinase K lyophilized 6 mg 30 mg Proteinase Buffer PB 1 8 mL 1 8 mL Collection Tubes 50 mL 4 25 Elution Tubes 0 5 mL 4 25 User manual 1 1 For preparation of working solutions and storage conditions see section 3 Composition of Elution Buffer BE 5 mM Tris HCl pH 8 5 4 MACHEREY NAGEL 06 2014 Rev 07 Genomic DNA from forensic samples 1 2 Reagents consumables and equipment to be supplied by user Reagents Ethanol 96 100 to prepare Buffer B5 and to adjust DNA binding conditions Consumables Disposable pipette tips 15mLand 50 mL centrifugation tubes Equipment Manual pipettors Centrifuge with swing out rotor suitable for 15 mL amd 50 mL tubes Suitable homogenization device e g mortar and pestle rotor stator Personal protection equipment e g lab coat gloves goggles 1 3 About this user manual It is strongly recommended that first time users of the NucleoSpin DNA Trace kit read the detailed protocol sections of this user manual Experienced u
5. Funken offener Flamme heiBen Oberfl chen fernhalten Keep container tightly closed Beh lter dicht verschlossen halten Avoid breathing dust Einatmen von Staub vermeiden Wear protective gloves eye protection Schutzhandschuhe Augenschutz tragen IF SWALLOWED Call a POISON CENTER doctor if you feel unwell BEI VERSCHLUCKEN Bei Unwohlsein GIFTINFORMATIONSZENTRUM Arzt anrufen IF ON SKIN Wash with plenty of water BEI KONTAKT MIT DER HAUT Mit viel Wasser waschen IF INHALED Remove victim to fresh air and keep at rest in a position com fortable for breathing BEI EINATMEN An die frische Luft bringen und in einer Position ruhigstellen die das Atmen erleichtert IF IN EYES Rinse continuously with water for several minutes Remove contact lenses if present and easy to do continue rinsing BEI KONTAKT MIT DEN AUGEN Einige Minuten lang behutsam mit Wasser sp len Vorhandene Kontaktlinsen nach M glichkeit entfernen Weiter sp len Call a POISON CENTER doctor if you feel unwell Bei Unwohlsein GIFTINFORMATIONSZENTRUM Arzt anrufen Rinse mouth Mund aussp len If skin irritation occurs Get medical advice attention Bei Hautreizung Arztlichen Rat einholen rztliche Hilfe hinzuziehen Get medical advice attention Bei anhaltender Hautreizung Arztlichen Rat einholen rztliche Hilfe hinzuziehen If experiencing respiratory symptoms Call a POISON CENTER doctor Bei S
6. 1 10 Die Substanz muss nicht als gef hrlich gekennzeichnet werden BW Guanidine hydrochloride Warning 226 302 210 233 280 36 50 isopropanol 319 336 301 312 20 50 96 305 351 338 330 Guanidinhydrochlorid 36 50 Isopropanol 20 50 Achtung 337 313 4034235 Proteinase K Proteinase K Iyophilized Proteinase K Iyophilisiert Danger 315 319 261 280 302 352 Gefahr 334 335 304 340 305 351 338 312 3324313 3374313 3424311 4034233 oe oe Hazard phrases H 226 Flammable liquid and vapour Fl ssigkeit und Dampf entz ndbar H 302 Harmful if swallowed Gesundheitssch dlich bei Verschlucken H 315 Causes skin irritation Verursacht Hautreizungen H 319 Causes serious eye irritation Verursacht schwere Augenreizung H 334 May cause allergy or asthma symptoms or breathing difficulties if inhaled Kann bei Einatmen Allergie asthmaartige Symptome oder Atembeschwerden verursachen H 335 May cause respiratory irritation Kann die Atemwege reizen H 336 May cause drowsiness or dizziness Kann Schl frigkit uud Benommenheit verursachen MACHEREY NAGEL 06 2014 Rev 07 9 Genomic DNA from forensic samples Precaution phrases P 210 P 233 P 261 P 280 P 301 312 P 302 352 P 304 340 P 305 351 338 P 312 P 330 P 332 313 P 337 313 P 342 311 P 403 233 P 403 235 Keep away from heat sparks open flames hot surfaces No smoking Von Hitze
7. 40922 100 mL Buffer B5 Concentrate 740921 20 for 100 mL Buffer B5 NucleoSpin DNA Trace 740943 25 1 set Bone Buffer Set Proteinase K 740506 100 mg 740505 50 50 mg DZ 740505 100 mg m ns Nuelaospine Fotensie Filiers 740988 10 50 250 10 50 250 pieces e or Eu m Forensic Filters 7 40988 50B 250B 1000B 50 250 1000 pieces Visit www mn net com for more detailed product information 6 3 Product use restriction warranty NucleoSpin DNA Trace kit components are intended developed designed and sold FOR RESEARCH PURPOSES ONLY except however any other function of the product being expressly described in original MACHEREY NAGEL product leaflets MACHEREY NAGEL products are intended for GENERAL LABORATORY USE ONLY MACHEREY NAGEL products are suited for QUALIFIED PERSONNEL ONLY MACHEREY NAGEL products shall in any event only be used wearing adequate PROTECTIVE CLOTHING For detailed information please refer to the respective Material Safety Data Sheet of the product MACHEREY NAGEL products shall exclusively be used in an ADEQUATE TEST ENVIRONMENT MACHEREY NAGEL does not assume any responsibility for damages due to improper application of our products in other fields of application Application on the human body is STRICTLY FORBIDDEN The respective user is liable for any and all damages resulting from such application 16 MACHEREY NAGEL 06 2014 Rev 07 Genomic DNA from forensic samples DNA RNA PROTEIN purificati
8. cids from large volumes because these columns are shaped like a funnel combining a large volume capacity with a small diameter of the binding membrane F means funnel The DNA isolated by NucleoSpin DNA Trace F Columns can be used directly for PCR or other enzymatic reactions Age storage conditions quantity and consistency of samples can affect DNA quality and therefore the protocol may be adapted accordingly e g increasing incubation time For successful DNA preparation it is essential that the sample is lysed well and separated afterwards only clear lysates should be loaded onto NucleoSpin DNA Trace F Columns in order to avoid clogging of the silica membrane The NucleoSpin DNA Trace kit allows purification of up to 20 ug of pure genomic DNA with an A A ratio of between 1 70 and 1 90 Some samples especially forensic samples may contain only traces of DNA However the amount will be sufficient for amplification and detection reactions Additional enzymes which are not included in the kit may be necessary for lysis of certain bacteria e g lysozyme lysostaphine Support protocol for the isolation of genomic DNA from human bones For this application additional Buffer T1 Buffer B3 and Proteinase K are necessary Therefore MACHEREY NAGEL offers the NucleoSpin DNA Trace Bone Buffer Set see ordering information This buffer set is especially designed for completion of the NucleoSpin DNA Trace kit It is
9. imes leads to shorter lysis times if no shaking water bath incubator is available Final incubation at 70 100 C for 5 min may be recommended for optimal denaturation and lysis of difficult samples e g dried old or clotted blood samples Clarify sample Afterwards any insoluble particles remaining in the sample have to be removed by centrifugation for 10 min at gt 5 000 x g in order to avoid clogging of the NucleoSpin DNA Trace membrane Adjust DNA binding conditions Add 3 5 mL ethanol 96 100 to 4mL cleared FLB lysate and vortex the mixture Use proportionally up scaled volumes of ethanol if more FLB lysate has been prepared in step 1 4 8 mL FLB 50 uL Proteinase K 56 C 1h 5 000 x g 10 min 3 5 mL ethanol Vortex MACHEREY NAGEL 06 2014 Rev 07 11 NucleoSpin DNA Trace 4 BindDNA Pipette mixture onto the NucleoSpin DNA Trace F Load sample Column 3 000 x g Centrifuge for 3 min at 3 000 x g Discard flow through 3 min with Collection Tube Put the NucleoSpin DNA Trace F Column into a fresh Collection Tube provided E 5 Wash silica membrane 1 wash 2 5 mL BW Add 2 5 mL Buffer BW to the NucleoSpin DNA Trace F 3 000 xg Column Centrifuge for 3 min at 3 000 x g 3 min 2 wash un Add 5 mL Buffer B5 to the NucleoSpin DNA Trace F 3 000 x g Column Centrifuge for 3 min at 3 000 x g discard flow 3 min through and reuse Collectio
10. mables during the production process is inactivated by means of the treatment with ethylene oxide in order to prevent the generation of accidental human profile by PCR amplification Ethylene oxide treatment has been shown to be the method of choice to prevent DNA profiles due to DNA contamination Shaw et al 2008 UV Gamma Electron beam Ethylene oxide 100 30 40 27 E 1396 87 MT MJ lj gt 30 70 Full profile Partial profile loadable ll Partial profile unloadable BB No profile Figure 1 According to Shaw et al 2008 Comparison of the effects of sterilization techniques on subsequent DNA profiling Int J Legal Med 122 29 33 MACHEREY NAGEL 06 2014 Rev 07 7 Genomic DNA from forensic samples 3 Storage conditions and preparation of working solutions Attention Buffers FLB and BW contain chaotropic salts Wear gloves and goggles CAUTION Buffers FLB and BW contain guanidine hydrochloride which can form highly reactive compounds when combined with bleach sodium hypochlorite DO NOT add bleach or acidic solutions directly to the sample preparation waste All kit components can be stored at room temperature 18 25 C and are stable for at least one year Upon storage especially at low temperatures a white precipitate may form in Lysis Buffer FLB Such precipitates have to be dissolved by incubating at 45 50 C for 10 min before use Before starting any NucleoSpin DNA Trace p
11. mple lysis Sample was not thoroughly homogenized and mixed with Buffer FLB Proteinase K The mixture has to be shaken continuously Alternatively prolong incubation time with Proteinase K No or poor Reagents not applied properly DNA yield Prepare Buffer B5 and Proteinase K solutions according to poor DNA instructions section 3 Add ethanol to lysates before loading quality them on NucleoSpin DNA Trace F Columns Suboptimal elution of DNA from the column Apply Elution Buffer BE 70 C directly onto the center of the silica membrane and incubate for 2 min Elution efficiencies decrease dramatically if elution is done with other buffers at pH s 7 0 RNA in sample Poor DNA f RNA free DNA is desired add 20 uL of RNase A solution qualty 20 mg mL to Lysis Buffer FLB and or suboptimal Carry over of ethanol Push aus Becertainto centrifuge 2 5 min at 3 000 x gin order to remove all DNA m M of ethanolic Buffer B5 before eluting the DNA If for any reason nz vimistie the level of Buffer B5 has reached the column outlet after the e dons second wash discard flow through Place the NucleoSpin DNA Trace F Column back into the Collection Tube and centrifuge again MACHEREY NAGEL 06 2014 Rev 07 15 Genomic DNA from forensic samples 6 2 Ordering information Product REF Pack of NucleoSpin DNA Trace 740942 4 25 4 25 preps NucleoSpin Funnel Column 740959 30 columns Buffer FLB 740322 500 500 mL Buffer BW 7
12. n Tube Cs 5mL B5 Era smaki Add 5 mL Buffer B5 to the NucleoSpin DNA Trace F 3 min Column Centrifuge for 3 min at 3 000 x g discard flow through and reuse Collection Tube 6 Dry silica membrane e 3 000 x g Centrifuge additional 10 min at 3 000 x g in order to 10 min remove Buffer B5 completely 7 Elute DNA Attach the supplied Elution Tube 0 5 mL with adaptor to the NucleoSpin DNA Trace F Column and insert 100 uL BE assembly into a new 50 mL centrifufe tube not provided 70 C Pipette 100 uL Buffer BE preheated to 70 C onto the NucleoSpin membrane and incubate for 2 min at room RT temperature 2 min Centrifuge for 3 min at 3 000 x g to collect the nucleic 3 000 x g acid containing fraction c 3 min Remove the elution tube containing the nucleic acids and keep it for further use 12 MACHEREY NAGEL 06 2014 Rev 07 NucleoSpin DNA Trace 5 2 Isolation of genomic DNA from human bones Before starting with the preparation please read remarks below Before starting with the preparation set incubators or water baths to 56 C and 70 C respectively Before elution equilibrate Elution Buffer BE to 70 C Attention The list numbers in this support protocol do not correspond with the list numbers in section 5 1 and protocol at a glance Additional Buffer T1 Buffer B3 and Proteinase K is necessary The NucleoSpin DNA Trace Bone Buffer Set REF 740943 25 is especially designed for completion of the Nucle
13. oSpin DNA Trace kit It is suited for 25 preparations of genomic DNA from human bones in conjunction with the NucleoSpin DNA Trace kit REF 740942 25 Preparation of Lysis Buffer B3 Transfer the total contents of Buffer B1 to Buffer B2 and mix well The resulting Buffer B3 is stable for at least one year at room temperature For each prep 2 mL additional buffer is necessary 0 5 M EDTA 0 25 M PO pH 8 not included in the NucleoSpin DNA Trace Bone Buffer Set 1 Prepare sample Mill 1 g bone to a fine powder 2 Pre lyse sample Add 2 mL buffer 0 5 M EDTA 0 25 M PO pH 8 and 7 mL Buffer T1 and 100 uL Proteinase K solution Vortex to mix Be sure that the samples are completely covered with lysis solution If processing several samples Proteinase K and Buffer T1 may be premixed directly before use Do never mix Bufer T1 and Proteinase K more than 10 15 min before addition to the sample Proteinase K tends to self digestion in Buffer T1 without substrate Incubate at 56 C overnight Afterwards incubate sample for 48 h at 4 C on a shaking incubator 3 Lyse sample Vortex the samples Add 8 mL Buffer B3 vortex vigorously and incubate at 70 C for 10 min Vortex briefly Centrifuge for 10 min at 5 000 x g and transfer the supernatant to a new microcentrifuge tube MACHEREY NAGEL 06 2014 Rev 07 13 NucleoSpin DNA Trace 4 Adjust DNA binding conditions Add 8 4 mL ethanol 96 100
14. on products of MACHEREY NAGEL are suitable for IN VITRO USES ONLY ONLY MACHEREY NAGEL products specially labeled as IVD are also suitable for IN VITRO diagnostic use Please pay attention to the package of the product IN VITRO diagnostic products are expressly marked as IVD on the packaging IF THERE IS NO IVD SIGN THE PRODUCT SHALL NOT BE SUITABLE FOR IN VITRO DIAGNOSTIC USE ALL OTHER PRODUCTS NOT LABELED AS IVD ARE NOT SUITED FOR ANY CLINICAL USE INCLUDING BUT NOT LIMITED TO DIAGNOSTIC THERAPEUTIC AND OR PROGNOSTIC USE No claim or representations is intended for its use to identify any specific organism or for clinical use included but not limited to diagnostic prognostic therapeutic or blood banking It is rather in the responsibility of the user or in any case of resale of the products in the responsibility of the reseller to inspect and assure the use of the DNA RNA protein purification products of MACHEREY NAGEL for a well defined and specific application MACHEREY NAGEL shall only be responsible for the product specifications and the performance range of MN products according to the specifications of in house quality control product documentation and marketing material This MACHEREY NAGEL product is shipped with documentation stating specifications and other technical information MACHEREY NAGEL warrants to meet the stated specifications MACHEREY NAGEL s sole obligation and the customer s sole remedy is limited
15. ordance with MACHEREY NAGEL 06 2014 Rev 07 17 Genomic DNA from forensic samples the stated specifications This warranty is exclusive and MACHEREY NAGEL makes no other warranty expressed or implied The warranty provided herein and the data specifications and descriptions of this MACHEREY NAGEL product appearing in MACHEREY NAGEL published catalogues and product literature are MACHEREY NAGEL s sole representations concerning the product and warranty No other statements or representations written or oral by MACHEREY NAGEL s employees agent or representatives except written statements signed by a duly authorized officer of MACHEREY NAGEL are authorized they should not be relied upon by the customer and are not a part of the contract of sale or of this warranty Product claims are subject to change Therefore please contact our Technical Service Team for the most up to date information on MACHEREY NAGEL products You may also contact your local distributor for general scientific information Applications mentioned in MACHEREY NAGEL literature are provided for informational purposes only MACHEREY NAGEL does not warrant that all applications have been tested in MACHEREY NAGEL laboratories using MACHEREY NAGEL products MACHEREY NAGEL does not warrant the correctness of any of those applications Last updated 07 2010 Rev 03 Please contact MACHEREY NAGEL GmbH amp Co KG Tel 49 24 21 969 270 tech bio 9 mn net com
16. rotocol prepare the following Wash Buffer B5 Add the indicated volume of ethanol 96 100 to Buffer B5 Concentrate Mark the label of the bottle to indicate that ethanol was added Store Wash Buffer B5 at room temperature 18 25 C for at least one year Before first use of the kit add the indicated volume see table below or on the bottle of Proteinase Buffer PB to dissolve lyophilized Proteinase K Proteinase K solution is stable at 20 C for at least 6 months NucleoSpin DNA Trace 4 preps 25 preps REF 740942 4 740942 25 Wash Buffer B5 12 mL 100 mL Concentrate Add 48 mL ethanol Add 400 mL ethanol Proteinase K 6 mg 30 mg Add 300 uL Add 1 5 mL Proteinase Buffer Proteinase Buffer 8 MACHEREY NAGEL 06 2014 Rev 07 Genomic DNA from forensic samples 4 Safety instructions The following components of the NucleoSpin DNA Trace kits contain hazardous contents Wear gloves and goggles and follow the safety instructions given in this section GHS classification Only harmful features do not need to be labeled with H and P phrases up to 125 mL or 125 g Mindergef hrliche Eigenschaften m ssen bis 125 mL oder 125 g nicht mit H und P S tzen gekennzeichnet werden Component Hazard contents GHS symbol Hazard Precaution phrases phrases Inhalt Gefahrstoff GHS Symbol H S tze P S tze FLB Guanidine hydrochloride Substance does not have to be specially labeled 1 10 as hazardous Guanidinhydrochlorid
17. sers however may refer to the Protocol at a glance instead The Protocol at a glance is designed to be used only as a supplemental tool for quick referencing while performing the purification procedure All technical literature is available on the internet at www mn net com Please contact Technical Service regarding information about changes of the current user manual compared to previous revisions 2 Product description 2 1 The basic principle NucleoSpin DNA Trace allows DNA isolation from cells tissue and many other sources Lysis is achieved by incubation of homogenized samples in a solution containing chaotropic ions and Proteinase K Appropriate conditions for binding of DNA to the silica membrane in the NucleoSpin DNA Trace F Columns are created by chaotropic salt and ethanol The binding process is reversible and specific to nucleic acids Contaminations are removed by repeated washing with 2 different ethanolic buffers Pure genomic DNA is finally eluted under low ionic strength conditions in a slightly alkaline elution buffer MACHEREY NAGEL 06 2014 Rev 07 5 Genomic DNA from forensic samples 2 2 Kit specifications NucleoSpin DNA Trace kit is designed for the preparation of highly pure genomic DNA from small amounts of any tissue cells and forensic samples for example dried blood spots The NucleoSpin DNA Trace F Columns included in the kit are ideally suited for collecting small amounts of nucleic a
18. suited for 25 preparations of genomic DNA from human bones in conjunction with the NucleoSpin DNA Trace kit REF 740942 25 Table 1 Kit specifications at a glance Parameter NucleoSpin DNA Trace Technology Silica membrane technology Format Funnel columns Sample material Forensic samples buccal swabs blood spots Sample size Forensic samples which can be extracted with up to 8 mL Lysis Buffer FLB in general 10 mg tissue 10 cells Fragment size 200 bp approx 50 kbp MACHEREY NAGEL 06 2014 Rev 07 Genomic DNA from forensic samples Table 1 Kit specifications at a glance Typical recovery gt 70 for amounts gt 10 ng Aveo Asa 1 7 1 9 Elution volume 100 uL Preparation time 60 min prep without Proteinase K incubation time which needs gt 1 h Binding capacity 20 ug Forensic quality product NucleoSpin DNA Trace is certified as forensic quality product Consumables used in forensics need to be treated carefully to prevent DNA contamination MACHEREY NAGEL therefore has a stringently controlled production process to avoid DNA contamination of consumables Further MACHEREY NAGEL uses ethylene oxide EO treatment to remove amplifiable DNA which might still be introduced during the manufacturing process MACHEREY NAGEL products carrying the forensic quality seal contain plastic materials that are EO treated This means DNA of any kind which might still be introduced into plastic consu
19. to replacement of products free of charge in the event products fail to perform as warranted Supplementary reference is made to the general business terms and conditions of MACHEREY NAGEL which are printed on the price list Please contact us if you wish to get an extra copy There is no warranty for and MACHEREY NAGEL is not liable for damages or defects arising in shipping and handling transport insurance for customers excluded or out of accident or improper or abnormal use of this product defects in products or components not manufactured by MACHEREY NAGEL or damages resulting from such non MACHEREY NAGEL components or products MACHEREY NAGEL makes no other warranty of any kind whatsoever and SPECIFICALLY DISCLAIMS AND EXCLUDES ALL OTHER WARRANTIES OF ANY KIND OR NATURE WHATSOEVER DIRECTLY OR INDIRECTLY EXPRESS OR IMPLIED INCLUDING WITHOUT LIMITATION AS TO THE SUITABILITY REPRODUCTIVITY DURABILITY FITNESS FOR A PARTICULAR PURPOSE OR USE MERCHANTABILITY CONDITION OR ANY OTHER MATTER WITH RESPECT TO MACHEREY NAGEL PRODUCTS In no event shall MACHEREY NAGEL be liable for claims for any other damages whether direct indirect incidental compensatory foreseeable consequential or special including but not limited to loss of use revenue or profit whether based upon warranty contract tort including negligence or strict liability arising in connection with the sale or the failure of MACHEREY NAGEL products to perform in acc
20. ymptomen der Atemwege GIFTINFORMATIONSZENTRUM Arzt anrufen Store in a well ventilated place Keep container tightly closed Beh lter dicht verschlossen an einem gut bel fteten Ort aufbewahren Store in a well ventilated place Keep cool K hl an einem gut bel fteten Ort aufbewahren For further information please see Material Safety Data Sheets www mn net com Weiterf hrende Informationen finden Sie in den Sicherheitsdatenbl ttern www mn net com 10 MACHEREY NAGEL 06 2014 Rev 07 NucleoSpin DNA Trace 5 Protocols 5 1 Isolation of genomic DNA from solid samples for example small amounts of cells or tissue forensic samples Before starting the preparation Check that Wash Buffer B5 was prepared according to section 3 Preheat Elution Buffer BE to 70 C Lyse sample Place the sample in a 15 mL centrifuge tube not provided and add 4 8 mL Buffer FLB The sample should be covered completely with Buffer FLB Solid samples should be homogenized by commercial tools pestle and mortar rotor stator homogeniser In general 10 mg tissue 10 cells or any DNA containing solid sample can be used Forensic samples dried blood spots chewing gum swabs etc should be covered completely with lysis buffer Add 50 uL Proteinase K stock solution mix by vortexing and incubate at 56 C in a shaking water bath until complete lysis is obtained 1 3 h or overnight Vortexing every 15 min 3 4 t
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