restriction enzyme to digest 16srRNA pcr product

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restriction enzyme to digest 16srRNA pcr product

by Kanchanadevi k :: Rate this Message:

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hi everyone

i need to digest 16srRNA pcr product with restrication enzyme
can any one tell me commonly used enzyme for 16srRNA pcr product.

actually i  have isolated around 50 strains of unknown bacterial sample
and i did 16srRNA amplication with universal primer 1492R, 27F
now i just wanted to see whether these bacterial strain are same or
different for further studies
for i wanted to digest them with restriction enzyme
so can any one tell me which enzyme i can use.


thank u so much




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K.Kanchana Devi,
Research Scholar,
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Parent Message unknown Re: restriction enzyme to digest 16srRNA pcr product

by Duncan Clark :: Rate this Message:

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Historians believe that in newspost
<mailman.714.1249327726.21502.microbio@...> on Tue, 4 Aug 2009,
Kanchanadevi k <kdevik@...> penned the following literary
masterpiece:
>i need to digest 16srRNA pcr product with restrication enzyme
>can any one tell me commonly used enzyme for 16srRNA pcr product.
>
>actually i  have isolated around 50 strains of unknown bacterial sample
>and i did 16srRNA amplication with universal primer 1492R, 27F
>now i just wanted to see whether these bacterial strain are same or
>different for further studies
>for i wanted to digest them with restriction enzyme
>so can any one tell me which enzyme i can use.

You will have to identify an enzyme that gives you enough fragments to
see if the isolates are similar or not. One cannot suggest a particular
enzyme as it will depend upon the sequence of your 16S. All I can
suggest is that you try a few 4bp recognition enzymes i.e. Hae III, Hpa
II, Sau3AI, Hinf I, Taq I, BstU I, Hha I etc. Pick the one that works
best for you.

Duncan
--
I love deadlines. I especially like the whooshing noise they make as
they go flying by.

Duncan Clark
GeneSys Ltd.
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