IGEM:IMPERIAL/2007/Projects/chassis/Testing/L1020

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L1020

E. coli S30 Extract System for Circular DNA

Contents:

  • 175µl Amino Acid Mixture Minus Cysteine, 1mM
  • 175µl Amino Acid Mixture Minus Methionine, 1mM
  • 175µl Amino Acid Mixture Minus Leucine, 1mM
  • 450µl S30 Extract, Circular (3 × 150µl)
  • 750µl S30 Premix Without Amino Acids
  • 250µl Luciferase Assay Reagent
  • 1ml Luciferase Dilution Reagent
  • 10µl pBESTluc DNA, Circular (1µg/µl)

Storage:

  • 70oC



Considerations:

  • Use highly purified DNA templates (e.g., CsCl- or gel-purified).
  • Inhibition of the expression caused by excess salts(NaCl ≥50mM), glycerol (≥1%), or by very small

amounts of magnesium or potassium salts.

  • In addition DNA purification kits should use sodium acetate rather than ammonium acetate. Typical yields range
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