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FRET 12-06-2006 12:58 PM

Question on FspI enzyme
 
Hi,
Does anyone have experience with FspI enzyme from NEB? This enzyme is common cutter for genomic DNA? the site of this enzyme in genome is quite less? I digested the DNA with this enzyme and it seems the resulting band are very large.
Does anyone have some idea on this?
Best wishes!
Thomas

admin 12-06-2006 09:25 PM

Re: Question on FspI enzyme
 
Hello again FRET!

yes FspI (NEB: [Only registered and activated users can see links. Click Here To Register...])
is commonly used to cut genomic DNA. An example is this paper here:
[Only registered and activated users can see links. Click Here To Register...]

Fragment sizes may vary from several kbs up...

the above mentioned paper in some cases also used EcoRI...

FRET 12-11-2006 08:11 AM

Re: Question on FspI enzyme
 
Thanks!
In this article, the FspI is used to cut the genomic DNA from E. coli. In human DNA, the CpGs are mostly under methylation. Thus. for human genomic and other mammalian genomic DNA, the Fsp I is not effective. I have tried and not detected any cut with southern blotting probes.
May this would help the other people!
Thomas

moleculardude 12-11-2006 11:05 PM

Re: Question on FspI enzyme
 
:)

Does this mean FspI would not cut CpG methylated restriction sites?

FRET 12-14-2006 10:47 AM

Re: Question on FspI enzyme
 
Maybe, I am not familar with this enzyme. I give up this and change to others! maybe this time works!

admin 12-14-2006 11:53 AM

Re: Question on FspI enzyme
 
I would pick the cheaper enzymes, they are not only cheap but usually very good!

BamHI is a good example :)


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