http://2010.igem.org/wiki/index.php?title=Team:Groningen/Protocols_for_Bacillus_subtilis_168&feed=atom&action=historyTeam:Groningen/Protocols for Bacillus subtilis 168 - Revision history2024-03-29T07:49:32ZRevision history for this page on the wikiMediaWiki 1.16.5http://2010.igem.org/wiki/index.php?title=Team:Groningen/Protocols_for_Bacillus_subtilis_168&diff=67757&oldid=prevNeima: New page: '''2-step transformation procedure for ''Bacillus subtilis'' 168''' *Cells are grown O/N at 37 °C in supplemented medium contains Spizizen's salts (without antibiotics) *Dilute c...2010-09-09T17:22:31Z<p>New page: '''2-step transformation procedure for ''Bacillus subtilis'' 168''' *Cells are grown O/N at 37 °C in <a href="/Supplemented_medium" title="Supplemented medium">supplemented medium</a> contains <a href="/Spizizen%27s_salts" title="Spizizen's salts">Spizizen's salts</a> (without antibiotics) *Dilute c...</p>
<p><b>New page</b></p><div><br />
'''2-step transformation procedure for ''Bacillus subtilis'' 168'''<br />
*Cells are grown O/N at 37 °C in [[supplemented medium]] contains [[Spizizen's salts]] (without antibiotics)<br />
*Dilute cells 10X in fresh medium (10 ml) and grow for 3 h at 37 °C, shaking <br />
*Make agar plates<br />
*Cells are then diluted 1:1 in [[starvation medium]](prewarmed, no antibiotic)<br />
*After 2 hours, approx. 1 μg DNA (BRB689; Cmr ''amyQ+'') is added to 100 μl cells in a 2 ml tube, and the mixture is incubated for 30 min at 37 °C (200 rpm)<br />
*Dilute 3-fold in prewarmed [[TY medium]] and plate after another 45 min (37 °C;200 rpm)<br />
*Dilutions (in starvation medium) for plating are:<br />
* 10^0 and 10^-1 on selective media<br />
* 10^-5 on non-selective plates (viable count)</div>Neima