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