Lactobacillus miniprep: Difference between revisions
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Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip. | Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip. | ||
*[[User:Kelly Drinkwater|Kelly Drinkwater]] 15:09, 17 August 2009 (EDT) Here's a [http://www1.qiagen.com/literature/render.aspx?id=467|user-generated protocol for B. subtilis] from the Qiagen website. It's the same as a normal miniprep except you add lysozyme with the buffer P1 to 1mg/mL and incubate at 37C for 10 min. There's nothing posted about how it performs. | |||
==References== | ==References== | ||
Latest revision as of 12:09, 17 August 2009
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Overview
A miniprep for Lactobacillus delbruckii (development in progress)
Procedure
- Incubate cells for 30-45 minutes in TES(50mM Tris[pH 7], 20mM EDTA, 0.2 mM sucrose) with 10mg/ml lysozyme and 70U/ml mutanolysin.
- Perform normal QIAGEN miniprep
- verify extraction of plasmid by sequencing or digesting and running on gel
Notes
Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!
- List troubleshooting tips here.
- You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
- Anecdotal observations that might be of use to others can also be posted here.
Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.
- Kelly Drinkwater 15:09, 17 August 2009 (EDT) Here's a protocol for B. subtilis from the Qiagen website. It's the same as a normal miniprep except you add lysozyme with the buffer P1 to 1mg/mL and incubate at 37C for 10 min. There's nothing posted about how it performs.
References
Relevant papers and books Serror et al, Applied and Environmental Microbiology, 2002
Contact
- Derek Ju (derekju [at] mit.edu)
or instead, discuss this protocol.
