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Team:Calgary/14 June 2010
From 2010.igem.org
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| + | [[Image:06.14.2010.R0040-E0430 Restreaks.png|thumb|right|400px|This was the restreak of Alex's, Patrick's, and Raida's three R0040-E0430 constructs. The colonies are slightly larger as they were left over the weekend.]] | ||
| + | [[Image:06.14.2010 R0040-E0430 (Amp) Gel.jpg|thumb|right|400px|This was the gel of the three R0040-E0430 constructs. It's not particularly clear, and it is possible that there are some errors with the protocol/materials.]] | ||
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'''Monday June 14, 2010''' | '''Monday June 14, 2010''' | ||
| - | Dev, Chris, and Jeremy: Today, we did the constructions of J13002 (a combination of a constitutive promoter with a RBS) and E0040 (GFP) and E0032 (YFP) to determine if the two fluorescent proteins were functional. As well, a plasmid switch was begun with the part J23032 into a Kan plasmid backbone so a selection pressure would be present when it was constructed with R0040. This construction was done using standard Biobrick cloning and then transformation into Top10 Competent cells. We received the parts from the registry in the form of agar stabs on friday June 11, 2010 but they were received too late to work with them. We also did a construction of B0015 to R0040 to begin the RNA detection circuit. The agar stabs were streaked onto plates and left to grow overnight of the parts I0500, K239000, K135000, and K274210. | + | ''Dev, Chris, and Jeremy:'' Today, we did the constructions of J13002 (a combination of a constitutive promoter with a RBS) and E0040 (GFP) and E0032 (YFP) to determine if the two fluorescent proteins were functional. As well, a plasmid switch was begun with the part J23032 into a Kan plasmid backbone so a selection pressure would be present when it was constructed with R0040. This construction was done using standard Biobrick cloning and then transformation into Top10 Competent cells. We received the parts from the registry in the form of agar stabs on friday June 11, 2010 but they were received too late to work with them. We also did a construction of B0015 to R0040 to begin the RNA detection circuit. The agar stabs were streaked onto plates and left to grow overnight of the parts I0500, K239000, K135000, and K274210. |
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| + | ''Alex, Patrick, and Raida:'' Surprisingly, we seemed to have picked good colonies because three out of four restreaks were distinctively yellow. The colonies are quite large, though, since we left them growing over the weekend. We did a colony PCR of these three restreaks, though the results were not very clear, nor very impressive. We will try and do another colony PCR when we finish switching and transforming the E0430 plasmid, which we also did today. We switched the E0430 (from colonies 3 and 4) to a pSB1K2 plasmid. Each was plated onto two kanamycin plates. | ||
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| + | As well, we also did an agar stab of the K239000 (DegP promoter) that arrived last Friday. This was streaked onto two ampicillin plates. It was discovered that the promoter was not in a normal BioBrick plasmid, but it is still likely that the BioBrick prefix and suffix are still in place, meaning we likely still able to digest the plasmid with EcoRI and PstI to switch the part into some other plasmid. Finally, we also transformed the E0420 (ECFP generator with no LVA tag) into cells. | ||
Revision as of 03:11, 18 June 2010
_Gel.jpg)
Monday June 14, 2010
Dev, Chris, and Jeremy: Today, we did the constructions of J13002 (a combination of a constitutive promoter with a RBS) and E0040 (GFP) and E0032 (YFP) to determine if the two fluorescent proteins were functional. As well, a plasmid switch was begun with the part J23032 into a Kan plasmid backbone so a selection pressure would be present when it was constructed with R0040. This construction was done using standard Biobrick cloning and then transformation into Top10 Competent cells. We received the parts from the registry in the form of agar stabs on friday June 11, 2010 but they were received too late to work with them. We also did a construction of B0015 to R0040 to begin the RNA detection circuit. The agar stabs were streaked onto plates and left to grow overnight of the parts I0500, K239000, K135000, and K274210.
Alex, Patrick, and Raida: Surprisingly, we seemed to have picked good colonies because three out of four restreaks were distinctively yellow. The colonies are quite large, though, since we left them growing over the weekend. We did a colony PCR of these three restreaks, though the results were not very clear, nor very impressive. We will try and do another colony PCR when we finish switching and transforming the E0430 plasmid, which we also did today. We switched the E0430 (from colonies 3 and 4) to a pSB1K2 plasmid. Each was plated onto two kanamycin plates.
As well, we also did an agar stab of the K239000 (DegP promoter) that arrived last Friday. This was streaked onto two ampicillin plates. It was discovered that the promoter was not in a normal BioBrick plasmid, but it is still likely that the BioBrick prefix and suffix are still in place, meaning we likely still able to digest the plasmid with EcoRI and PstI to switch the part into some other plasmid. Finally, we also transformed the E0420 (ECFP generator with no LVA tag) into cells.
