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Team:Baltimore/Notebook
From 2011.igem.org
| You can write a background of your team here. Give us a background of your team, the members, etc. Or tell us more about something of your choosing. | File:Baltimore logo.png 200px |
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Tell us more about your project. Give us background. Use this is the abstract of your project. Be descriptive but concise (1-2 paragraphs) | File:Baltimore team.png Your team picture |
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Notebook
Run through for abstract and scheduling: Start with plasmid with taq gene and pst1 gene Remove pst1 site from taq coding sequence • In order to remove the restriction site, do site directed mutagenesis (1day) • PCR • Digestion • Transformation • Screening (1day) o Dilute DNA o Colony PCR individually o Digestion of PCR product with pst1 o Run gel ~2500bp Add biobrick prefix/suffix to taq coding sequence • PCR and add the prefix and suffix as primers (one prefix has an extra AG-make sure to use the correct one) (1-2 days) o Run gel o Cut out of gel • Cut with restriction enzyme • Clean up DNA • Ligation with vector (could be vector with the terminator sequence, promoter and RBS) • Transformation Add a promoter, transcriptional terminator, ribosome binding site (RBS) • Screen colonies (1 day) o Colony PCR o Restriction Digestion o Clean up DNA • Sequence (1 day) Make taq protein Compare it to other enzymes and make sure it works
