High molecular DNA isolation for library construction
Michael L. Sullivan
mlsulliv at facstaff.wisc.edu
Fri Mar 31 09:28:54 EST 2000
Just this week I did a DNA prep from clover using Qiagen's DNeasy Plant
Maxi Kit. The kit was relatively inexpensive ($57 for 6 preps-- it more
than paid for itself in saving my time). I did a double prep using 1 g
tissue each, and ended up with about 150 ug of DNA. When I ran the some of
the DNA on a gel, it looked good, with fragments at least 20 kb. I haven't
used the DNA yet in a restriction digest or any other downstream
application, but I'm pretty confident it will work well.
Hope you find this helpful.
>I am going to isolate high molecular weight DNA from tomato leaves to
>construct library. In our lab, a former postDoc was trying to isolate
>protoplast for DNA extraction but it did not work well. Thus I am
>considering nuclei isolation method for DNA isolation. Moreover, referring
>to several papers, it seems the nuclei isolation method is better than
>protoplast isolation method.
>Does anybody have suggestion before I start this?
>Oregon State University
>Botany and Plant Pathology
>Cordely Hall 2082
>Corvallis, OR 97331-2902
Michael L. Sullivan, Ph.D
U.S. Dairy Forage Research Center
1925 Linden Drive West
Madison WI, 53706
(608) 264-5144 Phone
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