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Team:LMU-Munich/Lab Notebook/Protocols
From 2012.igem.org
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1. [https://static.igem.org/mediawiki/2012/5/56/LMU-Munich_2012_Media_for_Bacillus_subtilis.pdf Media and components for ''Bacillus subtilis''] | 1. [https://static.igem.org/mediawiki/2012/5/56/LMU-Munich_2012_Media_for_Bacillus_subtilis.pdf Media and components for ''Bacillus subtilis''] | ||
| + | This protocol gives the recipes for various media and the antibiotic concentrations used for ''B. subtilis''. | ||
2. [https://static.igem.org/mediawiki/2012/4/41/LMU-Munich_2012_Transformation_of_Bacillus_subtilis.pdf Transformation of ''Bacillus subtilis''] | 2. [https://static.igem.org/mediawiki/2012/4/41/LMU-Munich_2012_Transformation_of_Bacillus_subtilis.pdf Transformation of ''Bacillus subtilis''] | ||
| + | Protocol how to transform ''B. subtilis''. | ||
3. [https://static.igem.org/mediawiki/2012/0/00/LMU-Munich_2012_Isolation_of_chromosomal_DNA_from_Bacillus_subtilis_for_transformation.pdf Isolation of chromosomal DNA from ''Bacillus subtilis'' for transformation] | 3. [https://static.igem.org/mediawiki/2012/0/00/LMU-Munich_2012_Isolation_of_chromosomal_DNA_from_Bacillus_subtilis_for_transformation.pdf Isolation of chromosomal DNA from ''Bacillus subtilis'' for transformation] | ||
| + | A quick protocol how to isolate genomic DNA from ''B.subtilis'' to transform it to a different strain. By this mean you can very easily combine different B. subtilis genotypes. transformation of genomic DNA is much more efficient than plasmids. Isolated DNA with this protocol is not clean! | ||
4. [https://static.igem.org/mediawiki/2012/6/6b/LMU-Munich_2012_Isolation_of_genomic_DNA_from_Bacillus_%28for_PCR....%29.pdf Isolation of chromosomal DNA from ''Bacillus subtilis'' (for PCR....)] | 4. [https://static.igem.org/mediawiki/2012/6/6b/LMU-Munich_2012_Isolation_of_genomic_DNA_from_Bacillus_%28for_PCR....%29.pdf Isolation of chromosomal DNA from ''Bacillus subtilis'' (for PCR....)] | ||
| + | Isolation of pure genomic DNA from ''B. subtilis''. | ||
5. [https://static.igem.org/mediawiki/2012/7/7a/LMU-Munich_2012_Long-Flanking_Homology_PCR.pdf Long Flanking Homology PCR] | 5. [https://static.igem.org/mediawiki/2012/7/7a/LMU-Munich_2012_Long-Flanking_Homology_PCR.pdf Long Flanking Homology PCR] | ||
| + | PCR protocol for long flanking homology PCR. You might need those PCR products for interrupting genes with resistance cassettes. | ||
6. [https://static.igem.org/mediawiki/2012/6/6d/LMU-Munich_2012_Clean_deletions_in_Bacillus_subtilis.pdf Clean deletion of genomic fragments in ''Bacillus subtilis''] | 6. [https://static.igem.org/mediawiki/2012/6/6d/LMU-Munich_2012_Clean_deletions_in_Bacillus_subtilis.pdf Clean deletion of genomic fragments in ''Bacillus subtilis''] | ||
| + | Protocol for the production of markerless gene deletions. You will need the [http://www.ncbi.nlm.nih.gov/pubmed/15528558 pMAD] plasmid. This method is more time consuming than inserting resistnace cassettes but offers better cell perfomance. | ||
7. [https://static.igem.org/mediawiki/2012/6/66/LMU-Munich_2012_Spore_germination_assay.pdf Spore germination assay] | 7. [https://static.igem.org/mediawiki/2012/6/66/LMU-Munich_2012_Spore_germination_assay.pdf Spore germination assay] | ||
| + | Protocol to count efficiency of sporulation and germination. | ||
8. [https://static.igem.org/mediawiki/2012/7/70/LMU-Munich_2012_%C3%9F-Galactosidase_Assay_B._subtilis.pdf ß-Galactosidase Assay ''B. subtilis''] | 8. [https://static.igem.org/mediawiki/2012/7/70/LMU-Munich_2012_%C3%9F-Galactosidase_Assay_B._subtilis.pdf ß-Galactosidase Assay ''B. subtilis''] | ||
| - | + | Protocol how to measure ''lacZ'' gene activity in ''B. subtilis'' | |
Revision as of 14:48, 3 September 2012
The LMU-Munich team is exuberantly happy about the great success at the World Championship Jamboree in Boston. Our project Beadzillus finished 4th and won the prize for the "Best Wiki" (with Slovenia) and "Best New Application Project".
[ more news ]
Protocols
On this page, we will upload unique protocols to share with the public.
Since we work with Bacillus subtilis as chassis we focus on protocols how to work with that organism.
1. Media and components for Bacillus subtilis This protocol gives the recipes for various media and the antibiotic concentrations used for B. subtilis.
2. Transformation of Bacillus subtilis Protocol how to transform B. subtilis.
3. Isolation of chromosomal DNA from Bacillus subtilis for transformation A quick protocol how to isolate genomic DNA from B.subtilis to transform it to a different strain. By this mean you can very easily combine different B. subtilis genotypes. transformation of genomic DNA is much more efficient than plasmids. Isolated DNA with this protocol is not clean!
4. Isolation of chromosomal DNA from Bacillus subtilis (for PCR....) Isolation of pure genomic DNA from B. subtilis.
5. Long Flanking Homology PCR PCR protocol for long flanking homology PCR. You might need those PCR products for interrupting genes with resistance cassettes.
6. Clean deletion of genomic fragments in Bacillus subtilis Protocol for the production of markerless gene deletions. You will need the [http://www.ncbi.nlm.nih.gov/pubmed/15528558 pMAD] plasmid. This method is more time consuming than inserting resistnace cassettes but offers better cell perfomance.
7. Spore germination assay Protocol to count efficiency of sporulation and germination.
8. ß-Galactosidase Assay B. subtilis Protocol how to measure lacZ gene activity in B. subtilis
