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Team:Queens Canada/Notebook/Week17
From 2012.igem.org
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| - | <div id="protocolcontent">Protocol | + | <div id="protocolcontent"> |
| + | <table class="tableizer-table"> | ||
| + | <tr class="tableizer-firstrow"><th>Date</th><th>Protocol</th><th>People</th><th>DNA (if relevant)</th><th>Quantities and Parameters (if relevant)</th><th>Notes on Protocol</th></tr> <tr><td>19/08/2012</td><td>PCR overlap extension</td><td>Andrew</td><td>xylE XS and PP eCFP XS into nFF GFP + J04500</td><td> </td><td> </td></tr> <tr><td>19/08/2012</td><td>Digestion</td><td>Andrew</td><td>SmtA and Fmt with XS</td><td> </td><td> </td></tr> <tr><td>20/08/2012</td><td>Gel</td><td>David</td><td>DP fmt XS, DP SmtA XS, eCFP full construct PCR product</td><td> </td><td> </td></tr> <tr><td>20/08/2012</td><td>PCR overlap extension</td><td>Andrew</td><td> </td><td> </td><td> </td></tr> <tr><td>23/08/2012</td><td>Liquid cultures</td><td>Kevin</td><td>C. thermo x-dockerin II pet21b, CelD dock I pet1bb, C cell. Trx-dock pet32, C cell. cohesin pet32, C therm CipA Coh I pet21b, C therm CohII pQE30 , CipA Dockerin C therm pQE30,pCip7 (Coh7) C therm pQE30 ,CelD Dock C therm pQE30, C therm dock II pQE30</td><td> </td><td> </td></tr> <tr><td>24/08/2012</td><td>Minipreps</td><td>Kevin</td><td>see above</td><td> </td><td> </td></tr> <tr><td>27/08/2012</td><td>Heat shock transformation</td><td>Phillip</td><td>LinB pDE5TI7</td><td> </td><td> </td></tr> <tr><td>30/08/2012</td><td>Liquid cultures</td><td>Kevin</td><td>nFF GFP J04500 in DH5a, XL1 Blue, TOP10</td><td> </td><td> </td></tr> <tr><td>9/1/2012</td><td>PCR amplification of D3 domain</td><td>Kevin</td><td> </td><td> </td><td> </td></tr> <tr><td>01/09/2012</td><td>PCR</td><td>Kevin</td><td>Rv2579, Coh2, Doc2, xDoc2</td><td> </td><td> </td></tr> <tr><td>01/09/2012</td><td>Gel</td><td>Kevin</td><td>xylE XS</td><td> </td><td> </td></tr> <tr><td>01/09/2012</td><td>Gel</td><td>Kevin</td><td>D3, GE LinB, GE xylE XS</td><td> </td><td> </td></tr> <tr><td>03/09/2012</td><td>PCR OE</td><td>Kevin</td><td>LinB, CohII, Rv2579, D3</td><td> </td><td> </td></tr> <tr><td>03/09/2012</td><td>Digestion with DpnI</td><td>Phillip</td><td>LinB, CohII, Rv2579, D3</td><td> </td><td> </td></tr> <tr><td>03/09/2012</td><td>Miniprep</td><td>Kevin</td><td>linB, CohII, Rv2579, D3, DH5a + J04500</td><td> </td><td> </td></tr> <tr><td>05/09/2012</td><td>PCR of RFP</td><td>David</td><td> </td><td> </td><td> </td></tr> <tr><td>05/09/2012</td><td>Digestion with XP</td><td>Phillip</td><td>MPP LinB, rv2579, CohII, D3 full constructs, DH5a + J04500</td><td> </td><td> </td></tr> <tr><td>05/09/2012</td><td>PCR</td><td>Phillip</td><td>see above</td><td> </td><td> </td></tr> <tr><td>05/09/2012</td><td>PCR</td><td>Phillip</td><td>mgfp-5</td><td> </td><td> </td></tr> <tr><td>14/09/2012</td><td>Digestion with EP</td><td>Kevin</td><td>J04500, RFP FC J04500, CohII FC, D3 FC, Rv2579 FC, LinB FC</td><td> </td><td> </td></tr> <tr><td>15/09/2012</td><td>Liquid culture and miniprep</td><td>Kevin</td><td>xylE FC</td><td> </td><td> </td></tr> <tr><td>15/09/2012</td><td>Gel</td><td>Kevin</td><td>RFP, xylE FC, D GFP, D mgfp, D CohII, D xylE, D LinB, Rv2579</td><td> </td><td> </td></tr> <tr><td>15/09/2012</td><td>Ligation</td><td>Kevin</td><td>D3 domain into psb1c3</td><td> </td><td> </td></tr> <tr><td>16/09/2012</td><td>Gel</td><td>Beini</td><td>mgfp, xdoc, doc, xylE FC EP GE, D RFP GE, D mgfp, D cohII, D linB, D Rv2579</td><td> </td><td> </td></tr> <tr><td>16/09/2012</td><td>Digestion with SP</td><td>Beini</td><td>MPP DH5a + J04500, RBS B0034</td><td> </td><td>enzymatic purification using Biobasic kit</td></tr> <tr><td>16/09/2012</td><td>Liquid cultures and miniprep</td><td>Kevin, Phillip</td><td>D3 FC Ligation trials</td><td> </td><td> </td></tr> <tr><td>18/09/2012</td><td>Ligations</td><td>Kevin</td><td>xylE FC GE and RFP FC GE into pSB1C3</td><td> </td><td> </td></tr> <tr><td>18/09/2012</td><td>PCR overlap extension</td><td>Phillip</td><td>D RFP, D GFP, D mgfp, D cohII, D linB, D Rv2579 (all GEs)</td><td> </td><td> </td></tr> <tr><td>18/09/2012</td><td>Heat shock transformation of overlap extensions and ligations</td><td>Kevin</td><td>see above</td><td> </td><td> </td></tr> <tr><td>19/09/2012</td><td>Liquid cultures</td><td>Kevin</td><td>lig RFP, D3 RFP OE, D3 GFP OE, D3 linB OE, D3 Rv2579, D3 mgfp OE, D3 cohII OE, D3 ctrl</td><td> </td><td> </td></tr> <tr><td>21/09/2012</td><td>Minipreps</td><td>Aaron, Beini</td><td>see above</td><td> </td><td> </td></tr> <tr><td>22/09/2012</td><td>PCR OE of RFP with dockerin I</td><td>Victor</td><td>RFP, dockerin I</td><td>7 trials with varying ratios</td><td>used sec tag left primer and dock right primer</td></tr> <tr><td>22/09/2012</td><td>Heat shock transformation</td><td>Beini</td><td>xylE FC AK into XL1 Blue</td><td> </td><td> </td></tr> <tr><td>23/09/2012</td><td>Gel</td><td>Faisal</td><td>xylE, cohII, linB, Rv2579, mgfp, D3 ctrl, sxdoc, sdoc</td><td> </td><td> </td></tr> <tr><td>23/09/2012</td><td>PCR</td><td>Kevin</td><td>sdoc and XS RFP doc with sec tag 2 primer</td><td> </td><td> </td></tr> <tr><td>23/09/2012</td><td>Digestions</td><td>Beini</td><td>sxdoc XP, xylE with SN, xylE with EP, CohII with SP, mgfp with EP</td><td> </td><td> </td></tr> <tr><td>23/09/2012</td><td>Gel</td><td>Phillip</td><td>see above</td><td> </td><td> </td></tr> <tr><td>24/09/2012</td><td>Heat shock transformation</td><td>Kevin</td><td>J04450, RFP D3 OE</td><td> </td><td> </td></tr> <tr><td>24/09/2012</td><td>Glycerol stock</td><td>Kevin</td><td>xylE full fliC deletion construct in pSB1C3 in XL1 Blue</td><td> </td><td> </td></tr> <tr><td>24/09/2012</td><td>250mL culture</td><td>Kevin</td><td>xylE FC pSB1C3 XL1 Blue</td><td> </td><td> </td></tr> <tr><td>25/09/2012</td><td>Gel</td><td>Beini</td><td>gel extraction sxdoc RFP XP, sdoc RFP XP, xylE EP, MGFP D3 fliC EP</td><td> </td><td> </td></tr> <tr><td>25/09/2012</td><td>Heat shock transformation</td><td>Beini</td><td>MPP D3 FC lig into XL1 Blue, MPP D3 CohII OE into XL1 Blue</td><td> </td><td> </td></tr> <tr><td>25/09/2012</td><td>Ligations</td><td>Kevin</td><td>sdoc RFP J04500, sxdoc RFP J04500, mgfp pSB1C3</td><td> </td><td> </td></tr> <tr><td>26/09/2012</td><td>Liquid cultures</td><td>Beini</td><td>D3 FC, xylE FC AK, cohII FliC, mgfp5 FliC, sdoc RFP b0034, sxdoc RFP B0034</td><td> </td><td> </td></tr> <tr><td>26/09/2012</td><td>Miniprep</td><td>Phillip</td><td>see above</td><td> </td><td> </td></tr> <tr><td>29/09/2012</td><td>Heat shock transformations</td><td>Kevin</td><td>D3 FliC, D3 LinB FliC, D3 Rv2579 FliC, D3 Coh FliC, D3 RFP FliC, RFP deletion FliC, J04450, mgfp5 FliC</td><td> </td><td>transformed into TOP10</td></tr> <tr><td>30/09/2012</td><td>Liquid cultures</td><td>Beini</td><td>see above</td><td> </td><td> </td></tr> <tr><td>02/10/2012</td><td>Protein isolation</td><td>Kevin</td><td>mgfp5 FliC, J04450, D3 FliC, D3 LinB, RFP deletion FliC, D3 Rv2579 FliC, D3 RFP FliC, xylE deletion FliC, D3 CohII FliC</td><td>5 mL cutures</td><td>heated 65C for 15 mins</td></tr> <tr><td>02/10/2012</td><td>xylE assay</td><td>Kevin</td><td> </td><td> </td><td> </td></tr> <tr><td>03/10/2012</td><td>Flagella isolation precedure</td><td>Kevin</td><td> </td><td>successful pellet in all samples</td><td></td></tr></table> | ||
| + | </div> | ||
<div id="notebookcontent"> | <div id="notebookcontent"> | ||
<p style="font-size: 2em; text-align:middle'"> Given that the vast majority of our work now is in the lab, we will only be updating the "Labwork" section of the Notebook. </p> | <p style="font-size: 2em; text-align:middle'"> Given that the vast majority of our work now is in the lab, we will only be updating the "Labwork" section of the Notebook. </p> | ||
Revision as of 22:26, 26 October 2012
Control
Notebook - Week 17 and onwards
| Date | Protocol | People | DNA (if relevant) | Quantities and Parameters (if relevant) | Notes on Protocol |
|---|---|---|---|---|---|
| 19/08/2012 | PCR overlap extension | Andrew | xylE XS and PP eCFP XS into nFF GFP + J04500 | ||
| 19/08/2012 | Digestion | Andrew | SmtA and Fmt with XS | ||
| 20/08/2012 | Gel | David | DP fmt XS, DP SmtA XS, eCFP full construct PCR product | ||
| 20/08/2012 | PCR overlap extension | Andrew | |||
| 23/08/2012 | Liquid cultures | Kevin | C. thermo x-dockerin II pet21b, CelD dock I pet1bb, C cell. Trx-dock pet32, C cell. cohesin pet32, C therm CipA Coh I pet21b, C therm CohII pQE30 , CipA Dockerin C therm pQE30,pCip7 (Coh7) C therm pQE30 ,CelD Dock C therm pQE30, C therm dock II pQE30 | ||
| 24/08/2012 | Minipreps | Kevin | see above | ||
| 27/08/2012 | Heat shock transformation | Phillip | LinB pDE5TI7 | ||
| 30/08/2012 | Liquid cultures | Kevin | nFF GFP J04500 in DH5a, XL1 Blue, TOP10 | ||
| 9/1/2012 | PCR amplification of D3 domain | Kevin | |||
| 01/09/2012 | PCR | Kevin | Rv2579, Coh2, Doc2, xDoc2 | ||
| 01/09/2012 | Gel | Kevin | xylE XS | ||
| 01/09/2012 | Gel | Kevin | D3, GE LinB, GE xylE XS | ||
| 03/09/2012 | PCR OE | Kevin | LinB, CohII, Rv2579, D3 | ||
| 03/09/2012 | Digestion with DpnI | Phillip | LinB, CohII, Rv2579, D3 | ||
| 03/09/2012 | Miniprep | Kevin | linB, CohII, Rv2579, D3, DH5a + J04500 | ||
| 05/09/2012 | PCR of RFP | David | |||
| 05/09/2012 | Digestion with XP | Phillip | MPP LinB, rv2579, CohII, D3 full constructs, DH5a + J04500 | ||
| 05/09/2012 | PCR | Phillip | see above | ||
| 05/09/2012 | PCR | Phillip | mgfp-5 | ||
| 14/09/2012 | Digestion with EP | Kevin | J04500, RFP FC J04500, CohII FC, D3 FC, Rv2579 FC, LinB FC | ||
| 15/09/2012 | Liquid culture and miniprep | Kevin | xylE FC | ||
| 15/09/2012 | Gel | Kevin | RFP, xylE FC, D GFP, D mgfp, D CohII, D xylE, D LinB, Rv2579 | ||
| 15/09/2012 | Ligation | Kevin | D3 domain into psb1c3 | ||
| 16/09/2012 | Gel | Beini | mgfp, xdoc, doc, xylE FC EP GE, D RFP GE, D mgfp, D cohII, D linB, D Rv2579 | ||
| 16/09/2012 | Digestion with SP | Beini | MPP DH5a + J04500, RBS B0034 | enzymatic purification using Biobasic kit | |
| 16/09/2012 | Liquid cultures and miniprep | Kevin, Phillip | D3 FC Ligation trials | ||
| 18/09/2012 | Ligations | Kevin | xylE FC GE and RFP FC GE into pSB1C3 | ||
| 18/09/2012 | PCR overlap extension | Phillip | D RFP, D GFP, D mgfp, D cohII, D linB, D Rv2579 (all GEs) | ||
| 18/09/2012 | Heat shock transformation of overlap extensions and ligations | Kevin | see above | ||
| 19/09/2012 | Liquid cultures | Kevin | lig RFP, D3 RFP OE, D3 GFP OE, D3 linB OE, D3 Rv2579, D3 mgfp OE, D3 cohII OE, D3 ctrl | ||
| 21/09/2012 | Minipreps | Aaron, Beini | see above | ||
| 22/09/2012 | PCR OE of RFP with dockerin I | Victor | RFP, dockerin I | 7 trials with varying ratios | used sec tag left primer and dock right primer |
| 22/09/2012 | Heat shock transformation | Beini | xylE FC AK into XL1 Blue | ||
| 23/09/2012 | Gel | Faisal | xylE, cohII, linB, Rv2579, mgfp, D3 ctrl, sxdoc, sdoc | ||
| 23/09/2012 | PCR | Kevin | sdoc and XS RFP doc with sec tag 2 primer | ||
| 23/09/2012 | Digestions | Beini | sxdoc XP, xylE with SN, xylE with EP, CohII with SP, mgfp with EP | ||
| 23/09/2012 | Gel | Phillip | see above | ||
| 24/09/2012 | Heat shock transformation | Kevin | J04450, RFP D3 OE | ||
| 24/09/2012 | Glycerol stock | Kevin | xylE full fliC deletion construct in pSB1C3 in XL1 Blue | ||
| 24/09/2012 | 250mL culture | Kevin | xylE FC pSB1C3 XL1 Blue | ||
| 25/09/2012 | Gel | Beini | gel extraction sxdoc RFP XP, sdoc RFP XP, xylE EP, MGFP D3 fliC EP | ||
| 25/09/2012 | Heat shock transformation | Beini | MPP D3 FC lig into XL1 Blue, MPP D3 CohII OE into XL1 Blue | ||
| 25/09/2012 | Ligations | Kevin | sdoc RFP J04500, sxdoc RFP J04500, mgfp pSB1C3 | ||
| 26/09/2012 | Liquid cultures | Beini | D3 FC, xylE FC AK, cohII FliC, mgfp5 FliC, sdoc RFP b0034, sxdoc RFP B0034 | ||
| 26/09/2012 | Miniprep | Phillip | see above | ||
| 29/09/2012 | Heat shock transformations | Kevin | D3 FliC, D3 LinB FliC, D3 Rv2579 FliC, D3 Coh FliC, D3 RFP FliC, RFP deletion FliC, J04450, mgfp5 FliC | transformed into TOP10 | |
| 30/09/2012 | Liquid cultures | Beini | see above | ||
| 02/10/2012 | Protein isolation | Kevin | mgfp5 FliC, J04450, D3 FliC, D3 LinB, RFP deletion FliC, D3 Rv2579 FliC, D3 RFP FliC, xylE deletion FliC, D3 CohII FliC | 5 mL cutures | heated 65C for 15 mins |
| 02/10/2012 | xylE assay | Kevin | |||
| 03/10/2012 | Flagella isolation precedure | Kevin | successful pellet in all samples |
Given that the vast majority of our work now is in the lab, we will only be updating the "Labwork" section of the Notebook.
