What could be done to resuspend the mycobacterial pellet after chloroform:methanol wash so as to proceed for plasmid prep?

06 June 2015 1 6K Report

For plasmid DNA extraction from mycobacteria, cells have to be treated with chloroform:methanol to remove the mycolic acid layer which makes it very sturdy and hard? If not treated then have to compromise with the yield? Is there any other alternative?

Yoke Kqueen Cheah

Dear Minakshi,

Here are few references for your work:

http://www.ncbi.nlm.nih.gov/pubmed/20360362

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3698252/

http://www.einstein.yu.edu/tbresearch/Resources/Protocols/Protocol04.pdf

http://jcm.asm.org/content/43/5/2471.full

Regards

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