Difference between revisions of "Team:UMaryland/Notebook"
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First week review | First week review | ||
<ul> | <ul> | ||
| − | <li>Prepared a bacterial culture containing constitutive GFP (K584001) to grow overnights (E. coli in LB with chloramphenicol)</li> | + | <li>- Prepared a bacterial culture containing constitutive GFP (K584001) to grow overnights (E. coli in LB with chloramphenicol)</li> |
| − | <li>Performed a transformation in order to replicate our DNA with the constitutive GFP, and plated half of them on the plates containing chloramphenicol and used the other half for another overnight batch</li> | + | <li>- Performed a transformation in order to replicate our DNA with the constitutive GFP, and plated half of them on the plates containing chloramphenicol and used the other half for another overnight batch</li> |
| − | <li>Performed a mini-prep on our overnights with the constitutive GFP in order to isolate plasmids and verify that they were present through spectrophotometry<li> | + | <li>- Performed a mini-prep on our overnights with the constitutive GFP in order to isolate plasmids and verify that they were present through spectrophotometry<li> |
| − | <li>Started two separate 3A assemblies in order to piece together the pBAD promoter with the miraculin gene and the constitutive GFP gene to the SRNBC gene (homolog to Hok/Sok)</li> | + | <li>- Started two separate 3A assemblies in order to piece together the pBAD promoter with the miraculin gene and the constitutive GFP gene to the SRNBC gene (homolog to Hok/Sok)</li> |
| − | <li>Performed a transformation in order to clone our new plasmids, one containing the pBAD promoter and miraculin gene and the other containing SRNBC and the constitutive GFP</li> | + | <li>- Performed a transformation in order to clone our new plasmids, one containing the pBAD promoter and miraculin gene and the other containing SRNBC and the constitutive GFP</li> |
</ul> | </ul> | ||
<ul> | <ul> | ||
| − | <li>Transformed β-cyclase, AppY, and CRTBEY genes in bacteria to grow on plates overnight</li> | + | <li>- Transformed β-cyclase, AppY, and CRTBEY genes in bacteria to grow on plates overnight</li> |
| − | <li>Started a new 3A assembly for the SRNBC and constitutive GFP construct along with pBAD, constitutive colicinFy, and lysostaphin</li> | + | <li>- Started a new 3A assembly for the SRNBC and constitutive GFP construct along with pBAD, constitutive colicinFy, and lysostaphin</li> |
| − | <li>Final products were SRNBC + constitutive GFP, SRNBC + constitutive colicinFy, and pBAD + lysostaphin</li> | + | <li>- Final products were SRNBC + constitutive GFP, SRNBC + constitutive colicinFy, and pBAD + lysostaphin</li> |
| − | <li>Prepared overnights for the pBAD + miraculin construct that grew on the plates and the β-cyclase, AppY, and CRTBEY genes</li> | + | <li>- Prepared overnights for the pBAD + miraculin construct that grew on the plates and the β-cyclase, AppY, and CRTBEY genes</li> |
| − | <li>Performed minipreps on the overnights we had prepared the day before (for the β-cyclase, AppY, and CRTBEY genes) and shipped them off to be sequenced</li> | + | <li>- Performed minipreps on the overnights we had prepared the day before (for the β-cyclase, AppY, and CRTBEY genes) and shipped them off to be sequenced</li> |
| − | <li>Performed transformations on the SRNBC + constitutive GFP, SRNBC + constitutive colicinFy, and pBAD + lysostaphin digests and let them grow overnight</li> | + | <li>- Performed transformations on the SRNBC + constitutive GFP, SRNBC + constitutive colicinFy, and pBAD + lysostaphin digests and let them grow overnight</li> |
| Line 287: | Line 287: | ||
Miraculin | Miraculin | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- | + | <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>- 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> | ||
</ul> | </ul> | ||
<p style="font-size:24px;text-align:left;text-decoration: underline;"> | <p style="font-size:24px;text-align:left;text-decoration: underline;"> | ||
Designing gBlocks | Designing gBlocks | ||
<ul class="a"> | <ul class="a"> | ||
| − | <li>- Designed gblocks for epsilon-cyclase (from arabidopsis), pyocin and Hok/Sok</li> | + | <li>- Designed gblocks for epsilon-cyclase (from arabidopsis), pyocin and Hok/Sok, and ordered them</li> |
| + | <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> | ||
</ul> | </ul> | ||
<br> | <br> | ||
Revision as of 02:46, 19 September 2015
