Wolfgang Schechinger novalidaddress at
Thu Mar 30 16:03:55 EST 2006

Reads great.

I just wanted to add that, for most purposes, the Maniatis-Style
minipreps are even simplier:

Grow bacteria in LB in an eppi with some holes punched in the lid
Pellet bacteria by 5min at 5000g (approx)
add 150 ul of suspension buffer (you may use Merlin 1), scratch the
tube over an eppi stand for shockwave-redistribution, put on ice.
Add 150ul of Lysis Buffer (0.1M NaOH, 0.2% SDS, you may use Merlin 2),
scratch agin a few times, put on ice again.
Add 150ul of potassium acetate/acetic acid neutralization buffer (you
may use Merlin 3). Scratch again, spin 10-15 min at max. speed.
In order to precipitate the DNA, add the S/N to 500ul of isopropanol,
mix, spin for 15 min at max speed, remove S/N, wash again with 500 ul
of 70% ethanol which is stored at -20 in the dark. Spin 1 min at max
speed, remove S/N and allow to dry at RT.

I learned of this scratching method by hand_to_hand propaganda.

This protocol yields DNA suitable for digests, PCR and even sequencing
and transfections (at least for test purposes) at almost no cost and
shorter hands-on-time than almost any kit. Scale up to 5ml (to be grown
directly in 15 ml plastic tubes, precipitation is done in one or more
eppis) and more ml preps (you may need to increase buffer solutions to
300ul and morel) is possible.

I just wonder if the MMM (Magic Merlin Method) is used in these nice
{PCR, digest, etc} cleanup kits where you add binding buffer to your
sample, apply the mix on some white matter and wash with a wash buffer
where you have to add ethanol before use and then you elute with water
or TE.

So I wonder, too, if one might re-use these nice cleanup columns after

Anyone tried that yet?

Best regards,

Wolfgang Schechinger

Endocrine Research Lab,
BG & University Hospital Bergmannsheil

More information about the Methods mailing list