Difference between revisions of "Team:UMaryland/Notebook"
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body { | body { | ||
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#sidemenu { | #sidemenu { | ||
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Miraculin | Miraculin | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- RE digest on β-cyclase, AppY, CREB, and CRTBEY genes along with the pSB1C3 backbone and the pBAD + miraculin out of the | + | <li>- RE digest on β-cyclase, AppY, CREB, and CRTBEY genes along with the pSB1C3 backbone and the pBAD + miraculin out of the pSB1A3 backbone so that we could move the genes to the pSB1C3 backbone</li> |
<li>- Ran gel with all parts and cut out the bands</li> | <li>- Ran gel with all parts and cut out the bands</li> | ||
<li>- Mini-prepped SRNBC + constitutive colicinFy and shipped them off to be sequenced</li> | <li>- Mini-prepped SRNBC + constitutive colicinFy and shipped them off to be sequenced</li> | ||
<li>- Performed gel purification on gels from day before but they failed; they didn’t show up in the spectrophotometer</li> | <li>- Performed gel purification on gels from day before but they failed; they didn’t show up in the spectrophotometer</li> | ||
| − | <li>- Performed minipreps on pBAD+Miraculin in the | + | <li>- Performed minipreps on pBAD+Miraculin in the pSB1C3 backbone as well as the const. GFP+SRNBC</li> |
<li>- Performed RE digests on the previous mini-preps: EcoRI and PstI on pBAD + Miraculin, and XBa1 and Pst1 on SRNBC + Constitutive GFP</li> | <li>- Performed RE digests on the previous mini-preps: EcoRI and PstI on pBAD + Miraculin, and XBa1 and Pst1 on SRNBC + Constitutive GFP</li> | ||
</ul> | </ul> | ||
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<li>- Performed transformations of the constitutive GFP + SRNBC construct</li> | <li>- Performed transformations of the constitutive GFP + SRNBC construct</li> | ||
<li>- Transformations of SRNBC + Constitutive GFP failed</li> | <li>- Transformations of SRNBC + Constitutive GFP failed</li> | ||
| − | <li>- Performed PCR on the | + | <li>- Performed PCR on the pSB1C3 backbone in order to amplify it in preparation for a Gibson Assembly with the Hok/Sok gene</li> |
</ul> | </ul> | ||
<br> | <br> | ||
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Hok/Sok | Hok/Sok | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- Performed an RE digest on the | + | <li>- Performed an RE digest on the pSB1C3 backbone with XbaI and SpeI and then ran it through a gel to separate it out, to prepare for the moving of Hok/Sok into pSB1C3</li> |
</ul> | </ul> | ||
<br> | <br> | ||
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Hok/Sok | Hok/Sok | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- inserted Hok/Sok gblock into | + | <li>- inserted Hok/Sok gblock into pSB1C3 backbone using Gibson assembly</li> |
<li>- construct sent for sequencing </li> | <li>- construct sent for sequencing </li> | ||
</ul> | </ul> | ||
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Interlab | Interlab | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- performed a 3A assembly of each promoter + GFP in | + | <li>- performed a 3A assembly of each promoter + GFP in pSB1K3 (GFP in pSB1A3 and promoters in pSB1C3) </li> |
</ul> | </ul> | ||
<br> | <br> | ||
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<li>- const_promoter + RBS + RFP were replated from last week but produced no colonies </li> | <li>- const_promoter + RBS + RFP were replated from last week but produced no colonies </li> | ||
<li>- re-transformed original and new 3A assembly which produced the correct sequence </li> | <li>- re-transformed original and new 3A assembly which produced the correct sequence </li> | ||
| − | <li>- ligated const_promoter:QD-RFP in | + | <li>- ligated const_promoter:QD-RFP in pSB1C3 </li> |
| − | <li>- performed a 3A assembly of Hok/Sok plasmid with QD-RFP downstream in | + | <li>- performed a 3A assembly of Hok/Sok plasmid with QD-RFP downstream in pSB1K3 </li> |
<li>- site directed mutagenesis of quick degrading GFP (Bba_K750000) failed </li> | <li>- site directed mutagenesis of quick degrading GFP (Bba_K750000) failed </li> | ||
<ul class="a"> | <ul class="a"> | ||
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<ul class="a"> | <ul class="a"> | ||
<li>- 3A assembly of H/S + RFP failed </li> | <li>- 3A assembly of H/S + RFP failed </li> | ||
| − | <li>- ran a PCR of | + | <li>- ran a PCR of pSB1C3+H/S+RFP using 3 different rxn buffers </li> |
<li>- HF rxn buffer </li> | <li>- HF rxn buffer </li> | ||
<li>- GC rxn buffer </li> | <li>- GC rxn buffer </li> | ||
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<li>- ran PCR for GFP 1, GFP 3, and IL1 which produced the correct sized products </li> | <li>- ran PCR for GFP 1, GFP 3, and IL1 which produced the correct sized products </li> | ||
<li>- Gibson Assembly of GFP + IL1 produced colonies </li> | <li>- Gibson Assembly of GFP + IL1 produced colonies </li> | ||
| − | <li>- | + | <li>- pSB1C3 - IL3 did not have bands in gel of appropriate size </li> |
</ul> | </ul> | ||
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Lutein | Lutein | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- RE digests of pLAC-RFP in | + | <li>- RE digests of pLAC-RFP in pSB1C3 </li> |
<li>- ran a Gibson Assembly of E-cyclase/C3 and E-hydroxylase/C3 which produced colonies </li> | <li>- ran a Gibson Assembly of E-cyclase/C3 and E-hydroxylase/C3 which produced colonies </li> | ||
</ul> | </ul> | ||
Latest revision as of 23:57, 1 October 2015
