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DNA Isolation

Isolation and Quantification of Genomic DNA from Mycobacterium tuberculosis


The following protocol describes a "DNA mini-prep" procedure (Part A) for the isolation of chromosomal DNA from Mycobacterium tuberculosis and a procedure for quantifying the amount isolated (Part B).

Part A. Isolation of Nucleic Acids

NOTE: CAUTION! STEPS 1-10 SHOULD BE PERFORMED USING APPROPRIATE PROCEDURES FOR HANDLING MATERIAL POTENTIALLY CONTAMINATED WITH MYCOBACTERIUM TUBERCULOSIS. ALSO,THE LIDS OF THE EPPENDORF TUBES SHOULD BE CAREFULLY CLOSED AND OPENED SO AS TO AVOID SPLASHES AND AEROSOL =46ORMATION.


Part B. Determination of DNA Concentration


Appendix: Reagents/Solutions

Tween/albumin medium

Solution A

Solution B

5 M NaCl

CTAB/NaCl

Chloroform/Isoamyl alcohol

TE Buffer, pH 8.0

RNase

1X TNE Buffer



Written by Chet Cooper, Ph.D., January 14, 1993

Revised by Bob Jovell, M.S., December= 22, 1993


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