Upcoming lecturer: Dr. Renee Reijo-Pera

On Wednesday we will have the honor of having Dr. Renne Reijo-Pera as a guest lecturer in our class. Dr. Reijo Pera is visiting ONU as the Distinguished Keiser Lecturer of 2010.

Her lecture is titled Human Health, Development and Stem Cells, and it will be delivered on Monday, October 18, at 7:00 pm at the Freed Center

In our class Dr. Reijo-Pera will be talking about human preimplantation development and gene expression and pathways during the first few days of development. This topic is highly appropriate for our class and it will provide you with the opportunity to interact in an intimate setting with a world-class researcher. Take advantage of such opportunity!

Dr. Reijo-Pera is the Director of the Human Embryonic Stem Cell Research and Education Center at Stanford University. For more information see her lab web page, which includes information about her and her group, and her publications.

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Module 2, Lab 06 - Cloning - RED of the plasmid DNApurifications

(entry in progress)

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Module 2, Lab 06 - Cloning (GAPC gene from Arabidopsis and pJet1.2 plasmid)

Today we used the E. coli cultures you inoculated yesterday to do a small-scale plasmid DNA extraction ('miniprep') using Promega's Wizard® Plus SV Minipreps DNA Purification System.

Here's a recap of what we have done so far in the last couple of exercises:

Lab 5 - Nested PCR of the GAPC gene from Arabidopsis

Lab 6 - Cloning

• Ligation of PCR amplified GAPC gene onto the pJet1.2 plasmid
• Genetic transformation of E. coli with the pJet1.2 plasmid
• Cloning of genetically transformed E. coli
• Minipreps (purification of pJet1.2 plasmid)

Next, a restriction enzyme digestion to confirm successful plasmid DNA purification (lab 6) and setting sequencing reactions (lab 7)

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Module 3, Lab 08 - Genetic transformation of E. coli with the pGLO plasmid

Jen was kind enough to prepare new LB agar plates and redo the transformation of E. coli with the pGLO plasmid. Then she plated the transformed bacteria on the plates she prepared, some of which had ampicillin, arabinose, or both.

[Fantastic] Results are shown in the pictures below:

(entry in progress)

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Lecture, chapter 11 - RNA processing

tRNA processing

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Today we started chapter 11, on RNA processing, the fifth "stop" in our roadmap.

We mentioned the most basic processes through which RNA is modified (base modification, cleavage, and splicing) and described the more complex processes that are specific to eukaryotic RNA (5' capping and 3' polyadenylation).

We discussed in more detail the steps through which introns are spliced out the pre-mRNA molecule, and introduced the concept of alternative splicing.

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Lecture, chapter 10 - Gene regulation in eukaryotes

Today we covered chapter 10, on gene regulation in eukaryotes.

We highlighted the differences in between the regulatory processes in prokaryotes and the more complex processes in eukaryotes, mainly the facts that in eukaryotes many transcription factors are needed to aid transcription and that DNA is many times condensed in heterochromatin, and therefore unavailable for RNA polymerase and other proteins.

Among some of the processes that affect gene expression we discussed histone acetylation, a mechanism through which heterochromatin is relaxed into euchromatin, making DNA available for DNA binding proteins, and methylation, a mechanism through which genes can be repressed or silenced, in some cases because it promotes de-acetylation of histones and DNA condensation into heterochromatin.

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