CCLTM Complete Cell Lysis Solution169) - Code E203

Directions for Use

 

CCL is provided as a stable solution which contains 10 mg/ml Lysozyme (code 0663) and 10 mg/ml RNase A (code 0675). It can be substituted directly for lysozyme stock solutions in most alkaline lysis and boiling mini-preps. This solution eliminates RNA from your cell lysate, in addition to lysing your bacterial cells.

Below are two sample protocols for use of this solution in plasmid preps. Please remember that these are only sample protocols; if you have a protocol with which you are comfortable and familiar, this solution can be incorporated with only minor changes to your standard procedures. All volumes listed below are flexible; only the ratios need to be preserved.

 

Alkaline Lysis Method:
Spin down 1.5 ml of an overnight culture and remove the supernatant. Re-suspend pellet in 45 l of TE-glucose buffer (25mM Tris(code 0826) (pH 8.0), 1mM EDTA(code 0105) (pH 8.0), and 50mM glucose(code 0643)). Add 15 l of CCL and incubate for 15 minutes at 37C. Proceed as usual with your normal alkaline lysis protocol.

 

Boiling Lysis Method:
Spin down 1.5 ml of an overnight culture and remove supernatant. Resuspend pellet in 350 l STET solution(0.1M NaCl, 10mM Tris (pH 8.0), 1mM EDTA(code 0105) (pH 8.0), and 5% Triton X-100(code 0694)). Add 25 l CCL solution, and incubate at 37C for 15 minutes. Place tube in a boiling water bath for 40 seconds, and spin lysate for 10 minutes at room temperature. Proceed as usual with your normal boiling lysis method.

Expanded protocols are available upon request.

 

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