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== Notes == | == Notes == | ||
* We ligated all three inserts ( | * We ligated all three inserts (MFα2 promoter, BBa_K165057, and BBa_J63002) into the vector (BBa_K801000) using T4 DNA ligase. | ||
* We prepared and transformed ''E. coli'' with our ligations, then plated on LB+Amp plates. Positive (pUC) and negative (H<sub>2</sub> | * We prepared and transformed competent ''E. coli'' with our ligations, then plated on LB+Amp plates. Positive (pUC) and negative (H<sub>2</sub>O) transformation controls show the efficiency of the transformation and effectiveness of the selection, respectively. | ||
* Students came in on their own after this session to screen colonies with PCR. They then grew up and isolated promising construct pDNAs. | * Students came in on their own after this session to screen colonies with PCR. They then grew up and isolated promising construct pDNAs. | ||
Latest revision as of 23:08, 3 April 2013
Home Contact Internal Lab Members Projects Protocols Build-a-BUG Build-a-Gene 2016 Build-a-Gene 2015 Build-a-Gene 2014 Build-a-Gene 2013 Chalk Talks
Build-a-BUG: Series 2, Session 4
Saturday, March 23, 2013
Noon to 4:00 PM
This is the fourth Build-A-BUG workshop in a series of five on yeast. You will learn more about molecular cloning and continue to hone your basic lab techniques, while building a mating type detector. After troubleshooting an unexpected problem in Session 3, we will actually do ligations (linking DNA) and transformations of competent E. coli cells.
Background reading
Protocols
Materials
Notes
- We ligated all three inserts (MFα2 promoter, BBa_K165057, and BBa_J63002) into the vector (BBa_K801000) using T4 DNA ligase.
- We prepared and transformed competent E. coli with our ligations, then plated on LB+Amp plates. Positive (pUC) and negative (H2O) transformation controls show the efficiency of the transformation and effectiveness of the selection, respectively.
- Students came in on their own after this session to screen colonies with PCR. They then grew up and isolated promising construct pDNAs.
Whiteboard
Results
