RNA preparation for mycobacteria - (Nov/24/2005 )

Hi everybody, I need your help!!

I am looking for a protocol of RNA preparation for mycobacteria (H37Rv). But I have one problem normally I used Trizol method but this product cost a lot and our budget is empty for this year . So is anyone knows a cheap protocol to extract RNA????

Please helpppppppppppppppppppppppp me

Fred

Thanks a lot

Hi Biofred,

It's one of those things where you might find it cheapest to fork out a lot for the reagent because it will give you the best results, thus saving your time and wagecosts

Esh Mahenthiralingam in Parish and Stoker's Mycobacteria protocols, Humana Press (if you can afford £80, it's a good reference for any mycobacteriology lab) gives an older non-Trizol method using GITC and N-lauryl sarcosine buffer followed by P:CHCl3:IAA extract, and an ethanol ppt.

Hope this helps.

Hi,
You could try different "cheaper" methods:

1. Try the bead beating protocol, which involves to use of silicon beads and a GTC- Lysis buffer( check either Mycobacteria Protocols(Parish and Stoker) or Current Procols in Molecular Biology.

2.You could also try the classical "Camille Locht" method. Do a pubmed search for this author and you can get the protocol from any of his papers which has RNA extraction in them.

Good luck!

D.

Hi, there is another cheaper method, i. e. the simple Diethyl Pyrocarbonate (DEPC) method by William C. Summers (1970). The protocol is available online, and can be obtained by Google Search. It is simple and it works. However you still have to purchase the DNase to get rid of the DNA remnants in your RNA extract.
Good Luck.

Hi Fred
I scan the part of “Mycobacterial Protocol” book that has RNA extraction method. I don’t sure this method cheap for you.

enjoy
Yui

Hi Fred
I scan the part of “Mycobacterial Protocol” book that has RNA extraction method. I don’t sure this method cheap for you.

enjoy
Yui

Hi everybody

I am happy to see all this possibillity for my RNA extraction, thanks a lot for all you.

Have a nice day

Biofred

Isn't page 70 missing there?
Btw what we use is Trizol plus bead beater. Works very nice (including agilent quality control).