Team:WarrenCIndpls IN-HS/Notebook

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(Difference between revisions)
(May 26th: Primer Design)
(May 19th: Research on Parts)
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Promoter Region- site for RNA polymerase to attach to and begin transcription; yeast has many promoters within its genome that can be used to express metal detectors
Promoter Region- site for RNA polymerase to attach to and begin transcription; yeast has many promoters within its genome that can be used to express metal detectors
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Terminator Sequence- signals the end of transcriptoin to RNA polymerase
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Terminator Sequence- signals the end of transcription to RNA polymerase
Vector- Plasmid
Vector- Plasmid

Revision as of 13:04, 20 June 2011

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Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Safety Attributions


Contents

Notebook

May 4th: Bacterial and Yeast Transformation

Bacteria can be made to take in plasmids by heat shocking them; usually by icing them, puting them in a hot water bath, and then icing them again

May 11th: Gibson Assembly

Gibson Assembly pdf

May 19th: Research on Parts

Kozak Sequence- directs translation of mRNA for more efficiency and accuracy; the amount of protein synthesized from mRNA is dependent on the strength of the Kozak sequence

Promoter Region- site for RNA polymerase to attach to and begin transcription; yeast has many promoters within its genome that can be used to express metal detectors

Terminator Sequence- signals the end of transcription to RNA polymerase

Vector- Plasmid

    Multiple Cloning Site (MCS)- part of plasmid that can be cut open for genetic modification
    Origin of Replication (ORI)- sequence where replication is initiated
    Selection Markers
         -Ura3 is a selection marker for yeast
         -Ampicillin Resistance is a selection marker for bacteria based antibiotic resistance

May 26th: Primer Design

Primers- the 3' section must be complementary to the DNA template, the 5' end may have additional, non-complimentary base sequences to add restriction enzyme sites, and the 3' sections should not be complimentary to each other (increases risk of primer-dimers forming and inhibiting amplification); forward primers extend from start codon to stop codon while reverse primers work from stop codon to start codon

Our Primers:

1st Forward Primer contains an overhang to attach the biobrick to the plasmid, two restriction enzyme sites (for cutting the sequence out), and a primer for the construction of a new strand

2nd Forward Primer contains a primer for constructing a new strand to connect the translational unit to the biobrick

Reverse Primer contains an overhang to attach the biobrick to the multi-cloning site (on the plasmid), the other two restriction enzymes sites, and a primer for constructing a new strand