magic mp & sequenase

Cheung C. Yue ccy at po.CWRU.Edu
Fri Dec 4 09:44:39 EST 1992


My 0.02 cents worth on this subject.

1.  I am still in the dark ages, so I use 2 microgram of plasmid
    for my sequencing reactions (alkaline, EDTA, heat denatured),
    and I have had a frustrating time getting enough bacterial 
    growth to get the 10 microgram that Magic Miniprep was supposed
    to give me.  So I grew up a larger volume of bacteria (20 ml)
    culture, scaled up the lysis, and concentrated the DNA by a 
    single isopropanol precipitation, and took up the pellet in
    1 ml of Magicprep resin, and got the increased yield of plasmid
    that I want.  Nice supercoiled stuff as compared to a gel of
    the crude DNA which had a fair bit of nicked stuff (so supporting
    what Bruce Roe said).

2.  In doing all this mumble jumble, I also thought about not buying
    Promega's kit in the future.  They do sell the resin in bulk so
    I don't necessarily need to worry about the matrix.  What is nice about
    the kit are those spin "filters".  I vaguely recall seeing some
    supply company mentioning selling those things, but I can't recall
    which company.  Does anyone remember?

ccy at po.cwru.edu
-- 



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