Look in the article: Occurrences and stability of insertion sequences in Mycobacterium tuberculosis complex strains: evaluation of an insertion sequence-dependent DNA polymorphism as a tool in the epidemiology of tuberculosis.

I found this reasonably easily on google scholar and it was free :)

Hi

http://jcm.asm.org/content/32/10/2425.long

good luck

thanx margarete but i am confused because the paper states that "Chromosomal DNA was isolated as described by Ausubel et al".

I see the problem. Even Reza Hadavi's article refers to earlier articles which seem inaccessible. Sorry. Have you thought of trying other extraction methods? Particularly other phenol/chloroform extraction methods?

Here's an old one...

http://onlinelibrary.wiley.com/doi/10.1002/0471142727.mb0204s56/abstract;jsessionid=BB4FB953F22B461697F1F12EAD521A39.f03t02?deniedAccessCustomisedMessage=&userIsAuthenticated=false

hi

its better for you because this site not allow for uploading file.

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC270376/pdf/jcm00047-0232.pdf

Bacterial growth and chromosomal DNA isolation. Mycobacterial

strains were grown in 5 ml of Tween albumin

medium for 7 days to an optical density at 600 nm of 0.8 and

harvested by centrifugation (9). Chromosomal DNA was

isolated as described by Ausubel et al. (1). In short, 1.5 ml of

the concentrated culture was heated at 80°C for 20 min to kill

the cells. After centrifugation, the cells were resuspended in

500 ,ul of TE buffer (0.01 M Tris-HCl, 0.001 M EDTA [pH

8.0]). Lysozyme was added to a final concentration of 1

mg/ml, and the tube was incubated for i h at 37°C. Seventy

microliters of 10% sodium dodecyl sulfate (SDS) and 6 pil of

proteinase K (at a 10-mg/ml concentration) were added, and

the mixture was incubated for 10 min at 65°C. An 80-pul

volume of N-cetyl-N,N,N-trimethyl ammonium bromide

was added. The cups were vortexed briefly and incubated for

10 min at 65°C. An equal volume of chloroform-isoamyl

alcohol (24:1, vol/vol) was added, and the mixture was

vortexed for 10 s. After centrifugation for 5 min, 0.6 volume

of isopropanol was added to the supernatant to precipitate

the DNA. After 30 min at -20°C and centrifugation for 15

min, the pellet was washed once with 70% ethanol and the

air-dried pellet was redissolved in 20 pi of 0.1 x TE buffer

(0.001 M Tris-HCI, 0.0001 M EDTA [pH 8.0]).

good luck

Follw the attached article. Hope it'll work.

thak you Margarete, yes i tried chloroform extraction but it didnt work in my case. The paper which i am following states that " genomic DNA was isolated essentially by the method of Van soolingen (1995)". so I am now in search of the respective method, can i use ammonium chloride instead of ammonium bromide?

thank you all. It would be highly appreciated if anyone please answer my this query.

The paper which i am following states that " genomic DNA was isolated essentially by the method of Van soolingen (1995)". can i use ammonium chloride instead of ammonium bromide?

What is the stock concentration of N-Cetyl-N,N,N-trimethylammonium bromide used in this method?