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Question on FspI enzyme

Question on FspI enzyme - DNA Techniques

Question on FspI enzyme - Post questions and discuss DNA techniques and protocols such as DNA extraction, PCR, and the study of DNA-binding proteins.


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  #1  
Old 12-06-2006, 12:58 PM
Pipette Filler
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Default 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
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  #2  
Old 12-06-2006, 09:25 PM
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Default Re: Question on FspI enzyme

Hello again FRET!

yes FspI (NEB: http://www.neb.com/nebecomm/products/productR0135.asp)
is commonly used to cut genomic DNA. An example is this paper here:
http://nar.oxfordjournals.org/cgi/co...ull/26/24/5734

Fragment sizes may vary from several kbs up...

the above mentioned paper in some cases also used EcoRI...
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Old 12-11-2006, 08:11 AM
Pipette Filler
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Default 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
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Old 12-11-2006, 11:05 PM
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Default Re: Question on FspI enzyme



Does this mean FspI would not cut CpG methylated restriction sites?
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Old 12-14-2006, 10:47 AM
Pipette Filler
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Default 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!
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Old 12-14-2006, 11:53 AM
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Default 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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