BUGSS:Build-a-BUG:2-4: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
mNo edit summary |
mNo edit summary |
||
| (4 intermediate revisions by the same user not shown) | |||
| Line 7: | Line 7: | ||
''Noon to 4:00 PM''<br> | ''Noon to 4:00 PM''<br> | ||
This is the fourth Build-A-BUG workshop in a series of five | 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.<br> | ||
=== Background reading === | === Background reading === | ||
| Line 19: | Line 19: | ||
=== Materials === | === Materials === | ||
* [https://www.neb.com/products/m0202-t4-dna-ligase T4 DNA Ligase] | * [https://www.neb.com/products/m0202-t4-dna-ligase T4 DNA Ligase] | ||
== 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 (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. | |||
=== Whiteboard === | |||
[[Image:BAB2-4_Ligation.jpg|thumb|left|760px|alt=Diagram of desired construct after ligation|Diagram of desired construct after ligation]] | |||
[[Image:BAB2-4_Protocols.jpg|thumb|left|760px|alt=Ligation and transformation protocols|Ligation and transformation protocols]] | |||
=== Results === | |||
[[Image:BAB2-4_Transformants.jpg|thumb|left|760px|alt=Plates with transformants after 2 days|Plates with transformants after 2 days]] | |||
</div> | </div> | ||
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
