Team:Lethbridge/Notebook/Lab Work/July
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Back to [[Team:Lethbridge/Notebook|Notebook]]<br> | Back to [[Team:Lethbridge/Notebook|Notebook]]<br> | ||
Back to [[Team:Lethbridge/Notebook/Lab_Work|Lab Work]] | Back to [[Team:Lethbridge/Notebook/Lab_Work|Lab Work]] | ||
- | =July 2010= | + | =<font color="white">July 2010= |
- | ==July 3/2010== | + | ==<font color="white">July 3/2010== |
(In Lab: JS)<br> | (In Lab: JS)<br> | ||
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<b>Method:</b> Used [[Team:Lethbridge/Notebook/Protocols|Restriction of Plasmid DNA]] protocol and ligated the parts into pSB1C3.<br> | <b>Method:</b> Used [[Team:Lethbridge/Notebook/Protocols|Restriction of Plasmid DNA]] protocol and ligated the parts into pSB1C3.<br> | ||
- | ==July 5/2010== | + | ==<font color="white">July 5/2010== |
(In Lab: JV, AV, HB)<br> | (In Lab: JV, AV, HB)<br> | ||
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<font color ="Red">Picture to come!!!!!!!!!</font><br> | <font color ="Red">Picture to come!!!!!!!!!</font><br> | ||
- | ==July 5/2010 Evening== | + | ==<font color="white">July 5/2010 Evening== |
<b>Objective:</b> To over-express CFP complete in DH5α<br> | <b>Objective:</b> To over-express CFP complete in DH5α<br> | ||
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2) Went in the shaker at 6:50pm | 2) Went in the shaker at 6:50pm | ||
- | ==July 6/2010== | + | ==<font color="white">July 6/2010== |
(In lab: JV, AV, HB)<br> | (In lab: JV, AV, HB)<br> | ||
<b>Objective:</b> To continue the over-expression of CFP complete in DH5α<br> | <b>Objective:</b> To continue the over-expression of CFP complete in DH5α<br> | ||
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<font color ="Red">More to fill in, but Anthony does not understand the stuff written in the lab book</font><br> | <font color ="Red">More to fill in, but Anthony does not understand the stuff written in the lab book</font><br> | ||
- | ==July 8/2010== | + | ==<font color="white">July 8/2010== |
(In lab: JV, AV, HB, HS)<br> | (In lab: JV, AV, HB, HS)<br> | ||
<b>Objective:</b><br> | <b>Objective:</b><br> | ||
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<font color ="red"> Anthony does not understand the stuff written in the lab book.</font> | <font color ="red"> Anthony does not understand the stuff written in the lab book.</font> | ||
- | ==July 8/2010 - Evening== | + | ==<font color="white">July 8/2010 - Evening== |
(In lab: KG)<br> | (In lab: KG)<br> | ||
<b>Objective:</b>To transform TetR in pSB1A2 plasmid (BBa_C0040 - 2010 iGEM Distribution Kit Plate Well 4A) and pTetR in pSB1A2 plasmid (BBa_R0040 - 2010 iGEM Distribution Kit Plate Well 6) into DH5α.<br> | <b>Objective:</b>To transform TetR in pSB1A2 plasmid (BBa_C0040 - 2010 iGEM Distribution Kit Plate Well 4A) and pTetR in pSB1A2 plasmid (BBa_R0040 - 2010 iGEM Distribution Kit Plate Well 6) into DH5α.<br> | ||
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</table><br> | </table><br> | ||
- | ==July 9/2010== | + | ==<font color="white">July 9/2010== |
(In lab: JV)<br> | (In lab: JV)<br> | ||
<b>Objective:</b>To overexpress pLacI-mRBS-mms6-dT construct.<br> | <b>Objective:</b>To overexpress pLacI-mRBS-mms6-dT construct.<br> | ||
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<font color ="red">SDS PAGE picture!!!!!!!!!!!</font> | <font color ="red">SDS PAGE picture!!!!!!!!!!!</font> | ||
- | ==July 10/2010== | + | ==<font color="white">July 10/2010== |
(In lab: JV)<br> | (In lab: JV)<br> | ||
<b>Objective:</b>To determine if maxipreps finished on July 9th and 10th have significant concentrations of DNA.<br> | <b>Objective:</b>To determine if maxipreps finished on July 9th and 10th have significant concentrations of DNA.<br> | ||
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<font color ="red">AGAROSE GEL PICTURE</font> | <font color ="red">AGAROSE GEL PICTURE</font> | ||
- | ==July 12/2010== | + | ==<font color="white">July 12/2010== |
(In lab: AV,HB,JV)<br> | (In lab: AV,HB,JV)<br> | ||
<b>Objective:</b> Maxiprep pLacI (A2) from glycerol stock.<br> | <b>Objective:</b> Maxiprep pLacI (A2) from glycerol stock.<br> | ||
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4) Ligate restricted dT to the ends of the "Part 1" Biobricks.<br> | 4) Ligate restricted dT to the ends of the "Part 1" Biobricks.<br> | ||
- | ==July 12/2010 - Evening== | + | ==<font color="white">July 12/2010 - Evening== |
(In lab: KG,TF)<br> | (In lab: KG,TF)<br> | ||
<b>Objective:</b> Continue with addition of dT to the ends of mms6, xylE, and lumazine.<br> | <b>Objective:</b> Continue with addition of dT to the ends of mms6, xylE, and lumazine.<br> | ||
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</table><br> | </table><br> | ||
- | ==July 13/2010== | + | ==<font color="white">July 13/2010== |
(in lab: JV)<br> | (in lab: JV)<br> | ||
<b>Objective:</b> To determine if ligations of previous day (July 12/2010) were successful.<br> | <b>Objective:</b> To determine if ligations of previous day (July 12/2010) were successful.<br> | ||
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<font color ="red">AGAROSE GEL PICTURE</font><br> | <font color ="red">AGAROSE GEL PICTURE</font><br> | ||
- | ==July 14/2010== | + | ==<font color="white">July 14/2010== |
(in lab:J.S, K.G )<br> | (in lab:J.S, K.G )<br> | ||
<b>Objective:</b> Inoculate culture for maxi prep with placI | <b>Objective:</b> Inoculate culture for maxi prep with placI | ||
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Incubated reaction overnight at room temperature<br> | Incubated reaction overnight at room temperature<br> | ||
- | ==July 15/2010== | + | ==<font color="white">July 15/2010== |
(in lab: AV)<br> | (in lab: AV)<br> | ||
<b>Objective:</b> Maxiprep pLacI and mms6.<br> | <b>Objective:</b> Maxiprep pLacI and mms6.<br> | ||
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</table><br> | </table><br> | ||
- | ==July 15/2010 Evening== | + | ==<font color="white">July 15/2010 Evening== |
(in lab: AV)<br> | (in lab: AV)<br> | ||
<b>Objective:</b> transform ligations from July 14 and July 12,2010;xylE/dt, mms6/dt lumazine/dt into DH5&alpha. | <b>Objective:</b> transform ligations from July 14 and July 12,2010;xylE/dt, mms6/dt lumazine/dt into DH5&alpha. | ||
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<b>*</b>because of a shortage of plates not all transformations were plated at 50µL and 200µL | <b>*</b>because of a shortage of plates not all transformations were plated at 50µL and 200µL | ||
- | ==July 16,2010== | + | ==<font color="white">July 16,2010== |
(in lab: M.C, D.M)<br> | (in lab: M.C, D.M)<br> | ||
<b>Objective:</b> PCR amplify mms6-dT, xylE-dT, lumazine-dT legations along with dT maxi prep for comparison | <b>Objective:</b> PCR amplify mms6-dT, xylE-dT, lumazine-dT legations along with dT maxi prep for comparison | ||
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- | ==July 19, 2010== | + | ==<font color="white">July 19, 2010== |
(in lab: A.V, H.B)<br> | (in lab: A.V, H.B)<br> | ||
<b>Objective:</b> Purification pf pLacI and mms6 maxipreps done on July 14 and 16, 2010 using the biobasic protocol for purification of PCR products. | <b>Objective:</b> Purification pf pLacI and mms6 maxipreps done on July 14 and 16, 2010 using the biobasic protocol for purification of PCR products. | ||
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</table><br> | </table><br> | ||
- | ==July 19, 2010 Evening== | + | ==<font color="white">July 19, 2010 Evening== |
(K.G)<br> | (K.G)<br> | ||
<b>Objective:</b> Ligate together rbs-xylE and dT, lumazine and dT, also pLacI and sRBS.<br> | <b>Objective:</b> Ligate together rbs-xylE and dT, lumazine and dT, also pLacI and sRBS.<br> | ||
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<b>Results:</b> Lumazine Synthase with dT ligated onto it grew.<br> | <b>Results:</b> Lumazine Synthase with dT ligated onto it grew.<br> | ||
- | ==July 20, 2010== | + | ==<font color="white">July 20, 2010== |
(AV, HB)<br> | (AV, HB)<br> | ||
<b>Objective:</b>Miniprep lumazine-dt and 4 N-terminus tags and analyze.<br> | <b>Objective:</b>Miniprep lumazine-dt and 4 N-terminus tags and analyze.<br> | ||
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<font color ="red">AGAROSE GEL PICTURE</font><br> | <font color ="red">AGAROSE GEL PICTURE</font><br> | ||
- | ==July 20, 2010 Evening== | + | ==<font color="white">July 20, 2010 Evening== |
(AV, HB)<br> | (AV, HB)<br> | ||
<b>Objective:</b>Transform ligation done on July 19, 2010 in to competent DH5α cells.<br> | <b>Objective:</b>Transform ligation done on July 19, 2010 in to competent DH5α cells.<br> | ||
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*ECFP | *ECFP | ||
- | ==July 21, 2010== | + | ==<font color="white">July 21, 2010== |
(JV)<Br> | (JV)<Br> | ||
<b>Objective:</b> Isolate plasmid DNA from pTet, TetR, pET-28(a). | <b>Objective:</b> Isolate plasmid DNA from pTet, TetR, pET-28(a). | ||
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Ran SDS gel for 78 minutes at 200V<br> | Ran SDS gel for 78 minutes at 200V<br> | ||
- | ==July 21, 2010 Evening== | + | ==<font color="white">July 21, 2010 Evening== |
(TF, AS)<br> | (TF, AS)<br> | ||
<b>Objective:</b> Colony PCR to test for insertions of BioBrick construction, and for quality control of parts received from the Registry. Also to prepare DNA to be sent away for sequencing.<br> | <b>Objective:</b> Colony PCR to test for insertions of BioBrick construction, and for quality control of parts received from the Registry. Also to prepare DNA to be sent away for sequencing.<br> | ||
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<b>Results:</b> The PCR's did not work. | <b>Results:</b> The PCR's did not work. | ||
- | ==July 23, 2010== | + | ==<font color="white">July 23, 2010== |
(JV)<br> | (JV)<br> | ||
<b>Objective:</b> Insert xylE, Mms6, and lumazine synthase into pET-28(a).<br> | <b>Objective:</b> Insert xylE, Mms6, and lumazine synthase into pET-28(a).<br> | ||
<b>Method:</b> A restriction digest was performed on xylE and Mms6 and ran for 90 minutes at 37 C | <b>Method:</b> A restriction digest was performed on xylE and Mms6 and ran for 90 minutes at 37 C | ||
- | ==July 27, 2010== | + | ==<font color="white">July 27, 2010== |
(in lab: JV, AV, HB)<br> | (in lab: JV, AV, HB)<br> | ||
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<b>Results:</b> Qiagen Miniprep Kit produced a comparable quantity of DNA to [[Team:Lethbridge/Notebook/Protocols|Boiling Lysis Plasmid Preparation]]. Since the Qiagen Miniprep Kit is faster and easier, all minipreps will be done using the Qiagen Miniprep Kit. | <b>Results:</b> Qiagen Miniprep Kit produced a comparable quantity of DNA to [[Team:Lethbridge/Notebook/Protocols|Boiling Lysis Plasmid Preparation]]. Since the Qiagen Miniprep Kit is faster and easier, all minipreps will be done using the Qiagen Miniprep Kit. | ||
- | ==July 28, 2010== | + | ==<font color="white">July 28, 2010== |
(in lab: JV)<br> | (in lab: JV)<br> | ||
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<b>Method:</b> PCR amplified pSB1C3 received from iGEM headquarters. | <b>Method:</b> PCR amplified pSB1C3 received from iGEM headquarters. | ||
- | ==July 29, 2010== | + | ==<font color="white">July 29, 2010== |
(in lab: HB, AV)<br> | (in lab: HB, AV)<br> | ||
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<tr><td>Lumazine Synthase</td><td>2.66</td></tr></table> | <tr><td>Lumazine Synthase</td><td>2.66</td></tr></table> | ||
- | ==July 29, 2010 Evening== | + | ==<font color="white">July 29, 2010 Evening== |
(in lab: KG)<br> | (in lab: KG)<br> | ||