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
More information about the Methods