Labs 07 and 08

Friday October 09 2009

Lab 08 - Ligation and transformation

Students used bacterial colonies from the LB/Ampicillin agar plates incubated yesterday to inoculate LB/Ampicillin broth media. The broth media will be incubated for approx. 24 hours at 37ºC in a shaking water bath at approx. 200 rpm.

Lab 07 - Protein purification by chromatography

Students used the bacterial cultures inoculated yesterday to isolate Green Fluorescent Protein (GFP) by the process of Hydrophobic Interaction Chromatography (HIC).

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Labs 06, 07, and 08

Thursday October 08 2009

Lab 06 - pGLO bacterial transformation

Students analyzed the results of the bacterial cultures in LB, LB/Ampicillin, and LB/Ampicillin/Arabinose agar plates. A colony in the latter will be used to start lab 07.

Lab 07 - Protein purification by chromatography

Students used a colony from the LB/Ampicillin/Arabinose agar plates generated in lab 06 to inoculate an LB/Ampicillin/Arabinose broth tube. The broth was incubated at 37ºC for approx. 24 hours in a shaking water bath at approx. 200 rpm.

Lab 08 - Ligation and genetic transformation

Students prepared competent cells (E. coli) and transformed them with the pJet1.2 plasmid that has the GAPC gene (from Arabidopsis) insert.

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Lecture, chapter 7 - Proteins

We finished chapter 6, on transcription of genes, and went ahead and started chapter 7, on protein structure and function.

We talked about amino acids, the monomers that make up polypeptides, and how they are organized in four levels of structure. We also made the distinction between a polypeptide and a protein. We took a closer look to the primary and secondary structures of polypeptides (and proteins).

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Lecture, chapter 6 - Transcription of genes

Today we covered most of chapter 6, on transcription of genes.

We compared some details of transcription in prokaryotes and eukaryotes, including the kind of RNA polymerases involved in each one, the transcription factors involved in eukaryotic transcription, and some of the components of regulatory DNA.

Reminder: Next Wednesday (Oct 14) is the due date for the first draft of the review paper. The more advanced the paper the more feedback I'll be able to give you!

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