Team:Panama/Notebook

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====Notebook====
====Notebook====
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===Idea Project===
 
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Development of a genetically engineered ''E. coli'' that can produces the rhamnosyltranferase 1, for the production of a biosurfactant.
 
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Steps to follow:
 
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1.Find the gene sequences in pubmed.
 
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2.In which bacteria is found.
 
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3.After we found the gene, we designed two different set of primers.
 
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4.Look for the sequences of the restriction enzymes E, X, P and S, inside the gene sequence.
 
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5.If we found in the gene sequence a recognition sequence for the restriction enzymes.
 
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6. So we should look for a mutagenesis protocol or kit.
 
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7.We choose the Stratagene lightning mutagenesis kit. We also design three different set of primers, because the base pairs that surround the PstI unwanted sequence were different.
 
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===Meeting July 21===
 
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{{:Team:Panama/calendar2}}
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<center>
<gallery>
<gallery>
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Image:Panama-21-7-10-01.JPG|
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Image:INDICASAT-11.JPG|
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Image:Panama-21-7-10-02.JPG|
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Image:INDICASAT-05.JPG|
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Image:Panama-21-7-10-03.JPG|
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Image:INDICASAT-02.JPG|
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Image:Panama-21-7-10-05.JPG|
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Image:Panama-21-7-10-06.JPG|
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Image:Panama-21-7-10-07.JPG|
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Image:Panama-21-7-10-08.JPG|
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Image:DSC04818.JPG|
Image:DSC04818.JPG|
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</gallery>
 
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===Meeting June 26===
 
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At this moment we have two ideas to develop.  We are debating which idea is more feasible.
 
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Idea #1:  Carlos´s team idea
 
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We want to produce Rhamnolipid in bacteria (''E.coli'') to use it as a biosurfactant.
 
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We have to see how we can isolate the rhamnolipid and be sure that the translation is functional.
 
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<gallery>
 
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Image:ErnestoIdea.jpg|Ernesto's Idea
 
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</gallery>
 
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Steps to follow:
 
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1.Amplified the Rhamnosyltransferase  1  complex from ''Pseudomona auruginosa'' and clone this fragment. Size aprox 2.2 Kb.
 
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2.Ligation / Transformation
 
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3.Expression with reporter gen.
 
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4.Isolation
 
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5.Is the protein functional?
 
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6.Make sure that the rhamnolipid is produced in prescence of rhamnose and fatty acid.
 
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Notes:
 
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We can see the reaction (enzymatic measure) by spectrometry.
 
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See kind of ligation.
 
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But first, before the amplification we need to design our primers.
 
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Primers design:
 
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1.Check the gene sequence (In GenBank, FASTA format)
 
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2.Design the primer sequence.
 
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3.See kind of cloning.
 
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4.M13 tail for cleavage site.
 
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5.Look for cleavage site inside the rhamnosyltransferase 1 gene for our restriction enzymes.  We can´t cut our gene in the process.
 
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In summary the steps that we need to follow are:
 
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I.Primer design
 
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II.PCR or amplification
 
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III.Cloning
 
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IV.Expression
 
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V.Sequencing
 
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The most important steps are I and II. We need a good primers design and PCR if we want to have successful.
 
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Idea #2
 
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Ernesto team´s idea
 
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Production of Cecropin compound.  We want to produce the antibacterial cecropin compound. Naturally it is produced by insects and plants.
 
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We want to use ''Anopheles gambiae'', cecropin precursor. The gene is about 550 pb.
 
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The steps to follow are the same in both groups.
 
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Both groups have to design the primers, and have all the experimental design for this week.
 
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1.The sequence of the interested gene.
 
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2.Design the primers. (50-100pb more at the beginning and end of the sequence).
 
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3.Check for cloning sites. Which plasmid we are going to use, identify the restriction enzymes.
 
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4.Assemble the blocks. In paper assemble all the system. Promoter + Ribosomal binding site + interest gene + reporter gene + translation end site.
 
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All the design has to be on paper to analyze them and decided which idea is going to be the elected one.
 
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===Wetlab 103: Let's make an ''E. coli'' that fluoresce (or some simple BioBrick project)===
 
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* Instructor: INDICASAT, Dr. Carmenza Spadafora
 
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* Date:  (17h00-20h00 11th of June)
 
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* Venue:  INDICASAT
 
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* Description: We identify a simple project based on past iGEM work and do our first BioBrick protocol project. Nothing innovative, but an opportunity to practice the protocols.
 
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<gallery>
 
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Image:INDICASAT-01.JPG|INDICASAT
 
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Image:INDICASAT-02.JPG|INDICASAT
 
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Image:INDICASAT-03.JPG|INDICASAT
 
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Image:INDICASAT-04.JPG|INDICASAT
 
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Image:INDICASAT-05.JPG|INDICASAT
 
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Image:INDICASAT-06.JPG|INDICASAT
 
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Image:INDICASAT-07.JPG|INDICASAT
 
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Image:INDICASAT-08.JPG|INDICASAT
 
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Image:INDICASAT-09.JPG|INDICASAT
 
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Image:INDICASAT-10.JPG|INDICASAT
 
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Image:INDICASAT-11.JPG|INDICASAT
 
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Image:INDICASAT-12.JPG|INDICASAT
 
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</gallery>
 
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===Wetlab 102: Let's raise a few E. coli===
 
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* Instructor: Lorena Coronado and and Dr. Carmenza Spadafora
 
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* Date: (17h00-20h00 11th of June)
 
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* Venue: INDICASAT
 
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* Description: How does one handle ''E. col''i?
 
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<gallery>
 
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Image:INDICASAT_LECTURE-29.JPG
 
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Image:INDICASAT_LECTURE-30.JPG
 
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Image:INDICASAT_LECTURE-31.JPG
 
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Image:INDICASAT_LECTURE-32.JPG
 
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</gallery>
 
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===Wetlab 101: Tools of the lab and their use===
 
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* Instructor: Dr. Patricia Llanes
 
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* Date: (17h00-20h00 9th June)
 
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* Venue: INDICASAT
 
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* Description: How to handle pipettes, clean test tubes, etc.
 
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<gallery>
 
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Image:INDICASAT_LECTURE-19.JPG
 
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Image:INDICASAT_LECTURE-21.JPG
 
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Image:INDICASAT_LECTURE-22.JPG
 
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Image:INDICASAT_LECTURE-23.JPG
 
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Image:INDICASAT_LECTURE-24.JPG
 
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Image:INDICASAT_LECTURE-25.JPG
 
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Image:INDICASAT_LECTURE-26.JPG
 
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Image:INDICASAT_LECTURE-27.JPG
 
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Image:INDICASAT_LECTURE-28.JPG
 
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</gallery>
 
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===iGEM workshop follow up: Safety, ethics===
 
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* Instructor: Dr. Ricardo Lleonart
 
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* Date: (17h00-20h00 9th of June)
 
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* Venue: INDICASAT
 
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* Description: There is definetly a "safety considerations" requirement and we should address it early.
 
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<gallery>
 
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Image:INDICASAT_LECTURE-14.JPG|Dr. Ricardo Lleonart
 
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Image:INDICASAT_LECTURE-15.JPG|Dr. Ricardo Lleonart
 
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Image:INDICASAT_LECTURE-16.JPG
 
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Image:INDICASAT_LECTURE-17.JPG
 
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Image:INDICASAT_LECTURE-18.JPG
 
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</gallery>
 
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===iGEM workshop follow up: Software tools===
 
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* Instructor: Patrick / Sara
 
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* Date: (17h30-20h00 Monday 7th June)
 
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* Venue: INDICASAT
 
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* Description: Software tools available from the workshop.
 
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<gallery>
 
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Image:INDICASAT_LECTURE-13.JPG|Dra. Sara Pinzon
 
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</gallery>
 
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===DNA 104: BioBricks Protocol===
 
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* Instructor: Sara/Patrick
 
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* Date: (17h30-20h00 Monday 7th June)
 
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* Venue: INDICASAT
 
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* Description: Introduction to the BrioBricks protocol
 
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<gallery>
 
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Image:INDICASAT_LECTURE-09.JPG|Dr. Patrick Nee
 
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Image:INDICASAT_LECTURE-10.JPG
 
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Image:INDICASAT_LECTURE-11.JPG|Dr. Patrick Nee
 
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Image:INDICASAT_LECTURE-12.JPG|Dr. Patrick Nee
 
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</gallery>
 
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===INDICASAT lecture: Innovation===
 
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* Instructor: Dr. Jagannatha Rao, Director of INDICASAT
 
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* Date: (17h30-20h00 Friday 4th June)
 
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* Venue: INDICASAT
 
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* Description: Dr. Rao's lecture on how to innovate.
 
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<gallery>
 
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Image:INDICASAT_LECTURE-02.JPG
 
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Image:INDICASAT_LECTURE-04.JPG
 
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Image:INDICASAT_LECTURE-08.JPG
 
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</gallery>
 
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===INDICASAT lecture: Drug discovery in nature===
 
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* Instructor: Dr. Sergio Martinez
 
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* Date: (17h00-20h00 Thursday 3rd June)
 
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* Venue: INDICASAT
 
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* Description:Would be a brainstorming session as students start thinking about projects. It would be GREAT if we could take a molecule discovered by INDICASAT in coral/frogs/nature and put it in ''E. coli''!...
 
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===DNA 103: DNA modification, plasmids===
 
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* Instructor: Dr. Abby Guerra
 
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* Date: 26 May 17h00-20h00
 
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* Venue: UTP
 
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* Description:Introduction to long tested combinant DNA techniques, the role of plasmids in bacteria, and their use as a vector for DNA modification.
 
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<gallery>
 
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Image:Panama-05.JPG|Dr. Abby Guerra
 
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Image:Panama-06.JPG|Dr. Abby Guerra
 
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Image:Panama-07.JPG|Dr. Abby Guerra
 
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</gallery>
 
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===DNA 102: Protein creation relationship to cellular function. 26 May===
 
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* Instructor: Dr. Abby Guerra
 
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* Date: 26 May 17h00-20h00
 
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* Venue: UTP
 
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* Description: Introduction to how DNA drives cellular functions by creating proteins
 
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<gallery>
 
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Image:Panama-02.JPG|Dr. Abby Guerra
 
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Image:Panama-03.JPG|Dr. Abby Guerra
 
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</gallery>
 
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===DNA 101: Information store, replication. 26 May===
 
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* Instructor: Dr. Abby Guerra
 
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* Date: 26 May 17h00-20h00
 
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* Venue: UTP
 
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* Description: A basic refresher course on how DNA stores information in the cell, and how it is involved in cellular replication.
 
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<gallery>
 
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Image:Panama-01.JPG|Dr. Abby Guerra
 
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Image:Panama-04.JPG|Dr. Abby Guerra
 
</gallery>
</gallery>
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</center>

Latest revision as of 17:57, 14 October 2010

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Notebook

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