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restriction digestion


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#1 win

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Posted 15 September 2004 - 03:07 AM

I have done restriction digestion of plant genomic DNA with EcoRI and PstI and SacII but I was not able to get any bands .. the smear length aso was same. can someone suggest any thing regarding the smear paatern.

#2 sandrawrobel

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Posted 15 September 2004 - 06:20 AM

I would think that you will always see this kind of smear when genomic DNA of a large size (e. g. eukaryotic) is used.
There are just to many bands to differenciate one from the other.
Shouldn't you do some kind of Southern Blot in addition to the digestion or a PCR before to digest the product or should there really have been any differences in this stage already?

Sandra

#3 Nina

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Posted 15 September 2004 - 08:12 AM

Sometimes the gel results can have a smear pattern and in my case it was the wrong buffer in the agarose gel. When you make you gel with the agarose powder, you fill up with the same buffer that you use when you run the gel...

Maybe a long shoot...

/Nina




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