minipreps for sequencing

Eric C. Anderson anderson at pharmdec.wustl.edu
Tue Nov 1 15:03:16 EST 1994


In article <2AC3B02E01B938D9 at office.mmc.org>, varyc.mmcri at OFFICE.MMC.ORG
(Varyc) wrote:

> Opinionated netters,
> Are there any netters who are strongly opinionated as to the best protocol for large throughput reliable minipreps for cycle
> sequencing/ on ABI systems?, Or rating of prep systems for this use?

when we first bought our ABI sequencer, i went through a number of
different mini-prep systems to determine which one worked the best, gave
the greatest yield and was the easiest to use.  we settled on Wizard
mini-preps followed by an EtOH precipitation.  the preps i tested against
were Qiapreps which gave nice, clean DNA but the yield of a mini-prep was
usually too small to do more than one or two sequences from and after an
EtOH ppt. (which made them work even better) we were lucky to get one
sequence worth of template per prep; Bio-Rad's Prep-A-Gene which ABI
reccomends against using and i know why, the DNA quality was horrid and
yields were mediocre.  i also used alkaline lysis preps, with and without
PEG and/or EtOH precipitation.  the PEG/EtOH precipitated products were the
ones which worked as well in my hands as the Wizard or Qiaprep but took a
lot more time than either of those protocols, too much time to be worth it
for me.

just offering my 2 cents,
eric
-- 
*************************************
*Of course it's not true...         *
*But let's make the bastard deny it!*
*                        LBJ (1948) *
*************************************
eric c. anderson                                
anderson at pharmdec.wustl.edu
dept. of molecular bio. and pharm.               (314)362-3963 (lab)
washington univ. school of medicine              (314)862-2435 (home)
660 s. euclid box 8103                           (314)362-7058 (FAX)
st. louis, mo 63110



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