Wizard Plus SV Minipreps DNA Purification System Protocol

Wizard Plus SV Minipreps DNA Purification System Protocol -...

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Technical Bulletin Wizard ® Plus SV Minipreps DNA Purification System INSTRUCTIONS FOR USE OF PRODUCTS A1270, A1330, A1340, A1460 AND A1470. PRINTED IN USA. Revised 12/10 Part# TB225
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Promega Corporation · 2800 Woods Hollow Road · Madison, WI 53711-5399 USA Toll Free in USA 800-356-9526 · Phone 608-274-4330 · Fax 608-277-2516 · www.promega.com Printed in USA. Part# TB225 Revised 12/10 Page 1 1. Description .......................................................................................................... 1 2. Product Components and Storage Conditions ............................................ 3 3. Protocols .............................................................................................................. 4 A. Preparation of E. coli ............................................................................................ 4 B. Production of a Cleared Lysate. ......................................................................... 5 C. Plasmid DNA Isolation and Purification Protocols . ....................................... 5 4. Supplementary Information ............................................................................ 8 A. Selection and Preparation of Plasmids and E. coli Strains . ............................ 8 B. Choosing a Bacterial Strain. ................................................................................ 8 C. Use of Alkaline Protease . .................................................................................. 10 D. Considerations for Automated Fluorescent Sequencing. ............................. 10 5. Troubleshooting ............................................................................................... 11 6. Composition of Buffers and Solutions ....................................................... 14 7. References ......................................................................................................... 14 8. Related Products .............................................................................................. 15 1. Description The Wizard ® Plus SV Minipreps DNA Purification System (a,b) provides a simple, reliable method for rapid isolation of plasmid DNA (Figure 1). The entire miniprep procedure can be completed in 30 minutes or less, depending on the number of samples processed. This system can be used to isolate any plasmid from E. coli hosts but works most efficiently when the plasmid is less than 20,000bp in size. Purified plasmids can be used without further manipulation for automated fluorescent DNA sequencing as well as for other standard molecular biology techniques. When used for in vitro transcription reactions, the isolated plasmid DNA should be supplemented with a ribonuclease inhibitor such as Recombinant RNasin ® Ribonuclease Inhibitor (Cat.# N2511). The protocol presented in this Technical Bulletin is for isolation of plasmid DNA
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Wizard Plus SV Minipreps DNA Purification System Protocol -...

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