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

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:

• 70^oC

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