Team:LMU-Munich/Germination Stop

From 2012.igem.org

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==What Methods Did We Use to Knockout Germination Genes?==
==What Methods Did We Use to Knockout Germination Genes?==
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<font color="#000000"; size="2">Fig. 3: Long-flanking homology PCR as performed on gene ''cwlD'' to insert kanamycin-resistance (kan) cassette in its place. Fragments flanking ''cwlD'' are amplified. Up-reverse and down-forward primers have overhangs priming for kan cassette. Kan is simultaneously and separately amplified. Up and down ''clwD'' fragments and amplified kan fragments are put into PCR reaction. The result is a fragment containing ''cwlD'' up fragment, kan cassette, and ''cwlD'' down fragment. Entire fragment is put into reaction with genomic DNA of ''Bacillus''. Through recombination, new fragment is taken up in place of original fragment. Uptake is checked by growing cells on kanamycin.</font>
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<p align="justify">Two methods were employed to knock out germination: resistance cassette knockouts and clean deletions. Resistance cassette (RC) knockouts were performed using long-flanking-homology PCR (see [https://2012.igem.org/Team:LMU-Munich/Lab_Notebook/Protocols Protocols]). Single RC knockouts were created first; then they were combined to create multiple knockouts.</p>
<p align="justify">Two methods were employed to knock out germination: resistance cassette knockouts and clean deletions. Resistance cassette (RC) knockouts were performed using long-flanking-homology PCR (see [https://2012.igem.org/Team:LMU-Munich/Lab_Notebook/Protocols Protocols]). Single RC knockouts were created first; then they were combined to create multiple knockouts.</p>

Revision as of 13:20, 25 September 2012

iGEM Ludwig-Maximilians-Universität München Beadzillus

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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".

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