Related Manuals for Macherey-Nagel NucleoSpin 740727.10
Summary of Contents for Macherey-Nagel NucleoSpin 740727.10
- Page 1 Plasmid DNA purification User manual NucleoSpin ® Plasmid EasyPure August 2013 / Rev. 01...
- Page 2 50 μL Buffer AE RT, 1 min > 12,000 x g, 1 min MACHEREY-NAGEL GmbH & Co. KG · Neumann-Neander-Str. 6–8 · 52355 Düren · Germany Tel.: +49 24 21 969-270 · Fax: +49 24 21 969-199 · tech-bio@mn-net.com · www.mn-net.com...
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Page 3: Table Of Contents
Storage conditions and preparation of working solutions Safety instructions 4.1 Risk and safety phrases 4.2 GHS classification NucleoSpin Plasmid EasyPure protocol ® Appendix 6.1 Troubleshooting 6.2 Ordering information 6.3 References 6.4 Product use restriction / warranty MACHEREY-NAGEL – 08/ 2 013, Rev. 01... -
Page 4: Components
(dark blue rings) Collection Tubes (2 mL) Short protocol * For preparation of working solutions and storage conditions see section 3. **Composition of Elution Buffer AE: 5 mM Tris/HCl, pH 8.5 MACHEREY-NAGEL – 08 / 2 013, Rev. 01... -
Page 5: Reagents, Consumables, And Equipment To Be Supplied By User
All technical literature is available on the internet at www.mn-net.com. Please visit the MACHEREY-NAGEL website to verify that you are using the latest revision of this user manual. Please contact Technical Service regarding information about changes of the current user manual compared to previous revisions. -
Page 6: Product Description
DNA sequencing, PCR, or any kind of enzymatic manipulation. * For preparation of working solutions and storage conditions see section 3. ** Composition of Elution Buffer AE: 5 mM Tris/HCl, pH 8.5 MACHEREY-NAGEL – 08 / 2 013, Rev. 01... -
Page 7: Growth Of Bacterial Cultures
Since bacteria grow much faster without the burden of a high-copy plasmid, they take over the culture rapidly and the plasmid yield goes down regardless of the cell mass. Table 2 gives information on concentrations of commonly used antibiotics. * Hands-on-time MACHEREY-NAGEL – 08/ 2 013, Rev. 01... -
Page 8: Elution Procedures
About 90–100 % of bound nucleic acids can be eluted. * Maniatis T, Fritsch EF, Sambrook J: Molecular cloning. A laboratory manual, Cold Spring Harbor, Cold Spring, New York 1982. MACHEREY-NAGEL – 08 / 2 013, Rev. 01... - Page 9 8.0–8.5 since deionized water usually exhibits a pH below 7. Furthermore absorption of CO leads to a decrease in pH of unbuffered solutions. MACHEREY-NAGEL – 08/ 2 013, Rev. 01...
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Page 10: Storage Conditions And Preparation Of Working Solutions
10 preps 50 preps 250 preps 740727.10 740727.50 740727.250 6 mL 30 mL Wash Buffer AQ 6 mL (Concentrate) Add 24 mL ethanol Add 24 mL ethanol Add 120 mL ethanol MACHEREY-NAGEL – 08 / 2 013, Rev. 01... -
Page 11: Safety Instructions
** H azard labeling not neccessary if quantity per bottle below 125 g or mL (certificate of exemption according to 67/548/EEC Art. 25, 1999/45/EC Art. 12 and German GefStoffV § 20 (3) and TRGS 200 7.1). For further information see Material Safety Data Sheet. MACHEREY-NAGEL – 08/ 2 013, Rev. 01... -
Page 12: Ghs Classification
BEI VERSCHLUCKEN: Bei Unwohlsein GIFTINFORMATIONSZENTRUM oder Arzt anrufen. P 302+352 IF ON SKIN: Wash with plenty of soap and water. BEI KONTAKT MIT DER HAUT: Mit viel Wasser und Seife waschen. MACHEREY-NAGEL – 08 / 2 013, Rev. 01... - Page 13 Store in a corrosive resistant container with a resistant inner liner. In korrosionsbeständigem Behälter mit korrosionsbeständiger Auskleidung aufbewahren. For further information please see Material Safety Data Sheets (www.mn-net.com). Weiterführende Informationen finden Sie in den Sicherheitsdatenblättern (www.mn-net.com). MACHEREY-NAGEL – 08/ 2 013, Rev. 01...
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Page 14: Nucleospin ® Plasmid Easypure Protocol
Do not vortex to avoid shearing of genomic DNA! Clarification of lysate Centrifuge for 3 min at full speed (> 12,000 x g). > 12,000 x g, 3 min MACHEREY-NAGEL – 08 / 2 013, Rev. 01... - Page 15 > 12,000 x g, Centrifuge for 1 min at full speed (> 12,000 x g). 1 min Note: For more efficient elution procedures and alternative elution buffer (e.g., TE buffer or water) see section 2.4. MACHEREY-NAGEL – 08/ 2 013, Rev. 01...
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Page 16: Appendix
(rich media) at 37 ° C under shaking to avoid starvation and plasmid degradation. Incomplete neutralization • Mix thoroughly after addition of Buffer A3 until LyseControl has turned colorless without any traces of blue. MACHEREY-NAGEL – 08 / 2 013, Rev. 01... - Page 17 Buffer A2. Genomic DNA was sheared and thus liberated. quality Smeared plasmid bands on agarose gel • Especially when working with nuclease-rich strains, keep plasmid preparations on ice or frozen in order to avoid DNA degradation. MACHEREY-NAGEL – 08/ 2 013, Rev. 01...
- Page 18 DNA can be precipitated with ethanol and redissolved in Buffer AE or water. reactions Not enough DNA used for sequencing reaction • Quantify DNA by agarose gel electrophoresis before setting up sequencing reactions. MACHEREY-NAGEL – 08 / 2 013, Rev. 01...
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Page 19: Ordering Information
Birnboim, H.C., and J. Doly. 1979. A rapid alkaline extraction procedure for screening of recombinant plasmid DNA. Nucleic Acids Res. 7: 1513 - 1523. Vogelstein B., and D. Gillespie. 1979. Preparative and analytical purification of DNA from agarose. Proc. Natl. Acad. Sci. USA 76: 615 - 619. MACHEREY-NAGEL – 08/ 2 013, Rev. 01... -
Page 20: Product Use Restriction / Warranty
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;... - Page 21 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...