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DNA Kit Stability


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#1 MB1

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Posted 25 October 2012 - 03:18 AM

Dear All,

I have developed one DNA isolation kit for mycobacteria. this kit is based on Spin column principle.
Can anyone have an idea that how can i check the temperature based stability of my kit reagents. My kit contains buffers with salts and detergents, proteinase K and Lysozyme.

#2 bob1

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Posted 25 October 2012 - 12:26 PM

Store them at different temperatures for different periods of time, then run a standardized sample through the kit using the reagents that have been stored under the different conditions. You will also need to run normal condition controls, and probably do this sort of thing in bulk (e.g. do 5 separate samples for each condition) to ensure that you don't have sampling errors.

#3 MB1

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Posted 25 October 2012 - 09:50 PM

Thanks for the Reply,
I had desided to keep my kit at 4°C, Room Temperature and at 37°C.

For Proteinase K & Lysozyme: Though we know that these enzyme will be in active state to some extent at RT and 37°C, is it necessary to keep these Enzyme at such temperature? can I put them with remaining buffers?

For Samples: What kind of sample should I use? My kit can be use to isolate DNA from both, Gram positive as well as Gram negative cultures. Should I use fresh cultures of E.Coli & B. subtillis?


Thank you in advance
-MB1

#4 bob1

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Posted 25 October 2012 - 11:31 PM

You are wanting to know the stability of your kit - you need to test different storage conditions. Most people wouldn't store stuff at more than room temp for most kits -so I wouldn't worry about the 37 degree, but youmight want to consider whether you need to freeze the enzymes or whether they might be stable at fridge temp.

You know more about your kit than i do - you are the expert on what sort of samples would be appropriate.

#5 MB1

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Posted 26 October 2012 - 04:32 AM

Thank you.

I will revert back soon.




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