Size-Fractionation of DNA
William Alexander
ALEXANDERW at cber.cber.fda.gov
Wed Oct 18 05:33:22 EST 1995
Try using the Amicon microcon spin filters to get rid of SS and DS RNA/DNA
below 300 bases or 125 bp. There are different filters for different
molecular weight cut offs. Amicon also has a fax back sevice to send
protocols for desalting, size seperation, etc. (no affiliation) They may
cost a little but they are fast and you can avoid creating phenol waste,
which costs $ to dispose of.
Regards,
Bill Alexander
In Article <Pine.SUN.3.91.951018084008.3974A-100000 at chuma>,
leverone at CHUMA.CAS.USF.EDU ("Marianne Leverone ", BIO) wrote:
>Bob: I have used a CL-4B column to remove RNA from plasmid preps and the
>method worked well with high yields. I don't know about the size limits of
>this matrix. Look in a catalogue that sells the stuff and it will tell
>you what sizes of DNA that can be isolated. I always thought that it
>could not be used to size fractionate DNA, say for producing a library,
>because the plasmid isolation was based on the plasmid not getting
>trapped in the matrix but smaller, linear pieces did get trapped. How
>about it gang? Does anyone else know? Marianne Leverone, Biology Dept.,
>Univ. of South FLorida, Tampa, FL.
>
>On Mon, 16 Oct 1995 colvin at ouvaxa.cats.ohiou.edu wrote:
>
>> Before subclonning ds DNA generated by PCR into a plasmid, I would like to
>> remove small DNA fragments. The template used for PCR was a subtracted cDNA
>> library, so there are many different sized fragments present. One method
>> that I am aware of is to use a sepharose CL-4B column. Have others used this
>> method with success? Are there other methods that are simple, but still give
>> good recovery? Thank you.
>> Bob Colvin, Dept. Biological Sciences, Ohio Univ. Athens, OH, USA
>> colvin at ouvaxa.cats.ohiou.edu
>>
>>
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