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From 2011.igem.org

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Contents 1 Notebook 1.1 Calendar 1.1.1 July 1.1.2 August

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)
  • Dilute DNA
  • Colony PCR individually
  • Digestion of PCR product with pst1
  • 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)
  • Run gel
  • 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)
  • Colony PCR
  • Restriction Digestion
  • Clean up DNA
• Sequence (1 day)

Make taq protein

Compare it to other enzymes and make sure it works

Calendar

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July

Week 2: Tuesday, July 12; Wednesday, July 13; Thursday, July 14; Friday, July 15; Saturday, July 16; Sunday, July 17;

Week 3: Tuesday, July 19; Wednesday, July 20; Thursday, July 21; Friday, July 22; Saturday, July 23; Sunday, July 24;

Week 4: Tuesday, July 26; Wednesday, July 27; Thursday, July 28; Friday, July 29; Saturday, July 30; Sunday, July 31;

August

Week 1: Tuesday, August 2; Wednesday, August 3; Thursday, August 4; Friday, August 5; Saturday, August 6; Sunday, August 7;

Week 2: Tuesday, August 9; Wednesday, August 10; Thursday, August 11; Friday, August 12; Saturday, August 13; Sunday, August 14;

Week 3: Tuesday, August 16; Wednesday, August 17; Thursday, August 18; Friday, August 19; Saturday, August 20; Sunday, August 21;

Week 4: Tuesday, August 23; Wednesday, August 24; Thursday, August 25; Friday, August 26; Saturday, August 27; Sunday, August 28;