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Team:TU Delft/protocols/buffers stock solutions - Revision history
2024-03-29T08:44:23Z
Revision history for this page on the wiki
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http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/protocols/buffers_stock_solutions&diff=69347&oldid=prev
Ebrinkman at 18:45, 12 September 2010
2010-09-12T18:45:12Z
<p></p>
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<td colspan='2' style="background-color: white; color:black;">Revision as of 18:45, 12 September 2010</td>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline">=</del>=Buffers and Stock solutions<del class="diffchange diffchange-inline">=</del>=</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>=Buffers and Stock solutions=</div></td></tr>
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Ebrinkman
http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/protocols/buffers_stock_solutions&diff=69346&oldid=prev
Ebrinkman at 18:44, 12 September 2010
2010-09-12T18:44:19Z
<p></p>
<table style="background-color: white; color:black;">
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<td colspan='2' style="background-color: white; color:black;">Revision as of 18:44, 12 September 2010</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>'''TBE Buffer'''</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>'''<ins class="diffchange diffchange-inline">10x </ins>TBE Buffer'''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For 1 liter, dissolve in 800 mL H2O:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>For 1 liter, dissolve in 800 mL H2O:</div></td></tr>
</table>
Ebrinkman
http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/protocols/buffers_stock_solutions&diff=69345&oldid=prev
Ebrinkman at 18:43, 12 September 2010
2010-09-12T18:43:54Z
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<td colspan='2' style="background-color: white; color:black;">Revision as of 18:43, 12 September 2010</td>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''TBE Buffer'''</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>'''TBE Buffer'''</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>For 1 liter, dissolve in <del class="diffchange diffchange-inline">950 </del>mL H2O:</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>For 1 liter, dissolve in <ins class="diffchange diffchange-inline">800 </ins>mL H2O:</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div>Adjust to <del class="diffchange diffchange-inline">pH 8.0</del></div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div>Adjust <ins class="diffchange diffchange-inline">volume </ins>to <ins class="diffchange diffchange-inline">1 liter with H2O</ins></div></td></tr>
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Ebrinkman
http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/protocols/buffers_stock_solutions&diff=18384&oldid=prev
Ebrinkman at 17:56, 9 July 2010
2010-07-09T17:56:49Z
<p></p>
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<td colspan='2' style="background-color: white; color:black;">← Older revision</td>
<td colspan='2' style="background-color: white; color:black;">Revision as of 17:56, 9 July 2010</td>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>* Add agar in case a solid medium is required.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[</ins>*<ins class="diffchange diffchange-inline">]] </ins>Add agar in case a solid medium is required.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Autoclave on a liquid cycle. Once broth reaches room temperature, store in cold room.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Autoclave on a liquid cycle. Once broth reaches room temperature, store in cold room.</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|}</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>*Add agar in case a solid medium is required.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[</ins>*<ins class="diffchange diffchange-inline">]] </ins>Add agar in case a solid medium is required.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Once broth reaches room temperature, add to the broth in aseptic conditions the following amount of each solution:</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>Once broth reaches room temperature, add to the broth in aseptic conditions the following amount of each solution:</div></td></tr>
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<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|}</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>|}</div></td></tr>
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<tr><td class='diff-marker'>-</td><td style="background: #ffa; color:black; font-size: smaller;"><div><del class="diffchange diffchange-inline"> </del>*For M9-modified medium no glucose solution is added, instead 10 mL of filtered hydrocarbon/ hydrocarbon mixture are added per liter of medium. Be careful when adding the hydrocarbon/hydrocarbon mixture.</div></td><td class='diff-marker'>+</td><td style="background: #cfc; color:black; font-size: smaller;"><div><ins class="diffchange diffchange-inline">[[</ins>*<ins class="diffchange diffchange-inline">]] </ins>For M9-modified medium no glucose solution is added, instead 10 mL of filtered hydrocarbon/ hydrocarbon mixture are added per liter of medium. Be careful when adding the hydrocarbon/hydrocarbon mixture.</div></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"></td></tr>
<tr><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>NOTE: Autoclave the solutions 1 M MgSO4, 1M CaCl2, 20% glucose (w/v) (EXCEPT thiamine-HCl 1M) on a liquid cycle. Filter the solution Thiamine-HCl 1M using a 0.22 μm membrane, store the solution in a cold room when is not required.</div></td><td class='diff-marker'> </td><td style="background: #eee; color:black; font-size: smaller;"><div>NOTE: Autoclave the solutions 1 M MgSO4, 1M CaCl2, 20% glucose (w/v) (EXCEPT thiamine-HCl 1M) on a liquid cycle. Filter the solution Thiamine-HCl 1M using a 0.22 μm membrane, store the solution in a cold room when is not required.</div></td></tr>
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Ebrinkman
http://2010.igem.org/wiki/index.php?title=Team:TU_Delft/protocols/buffers_stock_solutions&diff=15734&oldid=prev
Ebrinkman: New page: ==Buffers and Stock solutions== '''Luria-bertani (LB)''' For 1 liter dissolve in H2O: {| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" |Compoun...
2010-07-05T14:07:18Z
<p>New page: ==Buffers and Stock solutions== '''Luria-bertani (LB)''' For 1 liter dissolve in H2O: {| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1" |Compoun...</p>
<p><b>New page</b></p><div>==Buffers and Stock solutions==<br />
<br />
<br />
'''Luria-bertani (LB)'''<br />
<br />
For 1 liter dissolve in H2O:<br />
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"<br />
|Compound/Ingredient <br />
|Amount required (1L of medium)<br />
|-<br />
|Tryptone<br />
|10 g<br />
|-<br />
|Yeast extract <br />
|5 g<br />
|-<br />
|NaCl<br />
|10 g<br />
|-<br />
|Agar+<br />
|15 g<br />
|}<br />
<br />
* Add agar in case a solid medium is required.<br />
<br />
Autoclave on a liquid cycle. Once broth reaches room temperature, store in cold room.<br />
<br />
<br />
'''M9'''<br />
<br />
For 1 liter dissolve in H2O:<br />
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"<br />
|Compound/Ingredient <br />
|Amount required (1L of medium)<br />
|-<br />
|Na2HPO4<br />
|6 g<br />
|-<br />
|KH2PO4<br />
|3 g<br />
|-<br />
|NaCl<br />
|0.5 g<br />
|-<br />
|(NH4)Cl<br />
|1 g<br />
|-<br />
|Agar*<br />
|15 g<br />
|}<br />
<br />
*Add agar in case a solid medium is required.<br />
<br />
Once broth reaches room temperature, add to the broth in aseptic conditions the following amount of each solution:<br />
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"<br />
|Solution <br />
|Amount(1L of medium)<br />
|-<br />
|1 M MgSO4<br />
|2 mL<br />
|-<br />
|1 M CaCl2<br />
|0.1 mL<br />
|-<br />
|20% Glucose (w/v)*<br />
|10 mL<br />
|-<br />
|1M Thiamine-HCl<br />
|1 mL<br />
|}<br />
<br />
*For M9-modified medium no glucose solution is added, instead 10 mL of filtered hydrocarbon/ hydrocarbon mixture are added per liter of medium. Be careful when adding the hydrocarbon/hydrocarbon mixture.<br />
<br />
NOTE: Autoclave the solutions 1 M MgSO4, 1M CaCl2, 20% glucose (w/v) (EXCEPT thiamine-HCl 1M) on a liquid cycle. Filter the solution Thiamine-HCl 1M using a 0.22 μm membrane, store the solution in a cold room when is not required.<br />
<br />
Store in cold room if the medium is not used.<br />
<br />
<br />
'''Antibiotics'''<br />
<br />
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"<br />
|Antibiotic <br />
|Final concentration in the medium<br />
|-<br />
|Ampicillin<br />
|100 μg/mL<br />
|-<br />
|Kanamycin <br />
|50 μg/mL<br />
|-<br />
|Chlorampenicol<br />
|35 μg/mL<br />
|-<br />
|Tetracycline<br />
|15 μg/mL<br />
|}<br />
<br />
All stock solutions must be kept at -20 ºC and used under aseptic conditions.<br />
<br />
<br />
'''TBE Buffer'''<br />
<br />
For 1 liter, dissolve in 950 mL H2O:<br />
<br />
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"<br />
|Compound/Ingredient <br />
|Amount required (1L of medium)<br />
|-<br />
|Tris<br />
|54 g<br />
|-<br />
|Boric Acid<br />
|27.5 g<br />
|-<br />
|0.5 M EDTA<br />
|4.65 g or 20 mL<br />
|}<br />
<br />
Adjust to pH 8.0</div>
Ebrinkman