Difference between revisions of "Team:Harvard/Notebook"
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| − | We designed an RBS library for csgG using the <a href="https://salislab.net/software/">Salis Lab RBS Library Calculator<a>, as well as the appropriate PCR primers to construct the library and ordered these sequences from IDT. | + | We designed an RBS library for csgG using the <a href="https://salislab.net/software/">Salis Lab RBS Library Calculator</a>, as well as the appropriate PCR primers to construct the library and ordered these sequences from IDT. |
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| − | <div class="panel-body">Our DNA parts ordered the previous week arrived and we conducted PCRs with the primers we designed and a plasmid containing the sequences for csgA, csgB, csgC, csgE, csgF, and csgG obtained from the Joshi Lab to modify the RBS sequence in front of csgG. After verifying our PCR products with a gel, we used Gibson Assembly to put our cloned parts in an expression vector. </div> | + | <div class="panel-body"> |
| + | <h5>Day 1</h5> | ||
| + | <br> | ||
| + | Our DNA parts ordered the previous week arrived and we conducted PCRs with the primers we designed and a plasmid containing the sequences for csgA, csgB, csgC, csgE, csgF, and csgG obtained from the Joshi Lab to modify the RBS sequence in front of csgG. | ||
| + | <br><br> | ||
| + | <h5>Day 2</h5> | ||
| + | <br> | ||
| + | After verifying our PCR products with a gel, we used Gibson Assembly to put our cloned parts in an expression vector with kanamycin resistance. We then transformed our newly formed plasmids into competent cells using heat shock and plated them on agar plates with kanamycin. | ||
| + | <br><br> | ||
| + | <h5>Day 3</h5> | ||
| + | <br> | ||
| + | After leaving our plates in an incubator overnight, we picked single colonies and cultured them in 5 mL falcon tubes with liquid LB and kanamycin. | ||
| + | |||
| + | </div> | ||
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</div> | </div> | ||
Revision as of 01:00, 1 November 2017
Lab Notebook
We designed an RBS library for csgG using the Salis Lab RBS Library Calculator, as well as the appropriate PCR primers to construct the library and ordered these sequences from IDT.
Day 1
Our DNA parts ordered the previous week arrived and we conducted PCRs with the primers we designed and a plasmid containing the sequences for csgA, csgB, csgC, csgE, csgF, and csgG obtained from the Joshi Lab to modify the RBS sequence in front of csgG.
Day 2
After verifying our PCR products with a gel, we used Gibson Assembly to put our cloned parts in an expression vector with kanamycin resistance. We then transformed our newly formed plasmids into competent cells using heat shock and plated them on agar plates with kanamycin.
Day 3
After leaving our plates in an incubator overnight, we picked single colonies and cultured them in 5 mL falcon tubes with liquid LB and kanamycin.
No lab work :(
