Re Protocol for Turbo-lifting/screening

Martin Leach leach at mbcrr.harvard.edu
Fri Sep 24 06:18:53 EST 1993


Hi Netters,

Received this protocol from mkennedy at chmeds.ac.nz and is taken from NAR
17:452 by Buluwela et al.

Martin Kennedy writes:

>I use an even easier method, "turbo lifting" by (Above ref.). Do your lifts >onto filters, place them onto 3MM paper soaked in "turbo juice" (2xssc, 5%sds), >and microwave the whole lot for 2.5 minutes. You can optionally UV cross-link >but its not necessary.

Haven't tried it yet but it sounds good.



I also have another quick method for purifying bands from agarose gels:

1.Excise band, trimming agarose.

2.Prepare silanized glass-wool. (Add dimethyldichlorosilane solution to
glass wool, shake a few times, rinse thorougly with distilled water, store
at RT in distilled water indefinetly.)

3.Cut caps off two 1.5 ml tubes and one 0.6 ml tube. Make hole in base of
0.6 ml tube with 23g syringe needle. Cut a small amount of silanized glass
wool and pack in bottom of 0.6 ml tube (3-4mm in depth). Place 0.6ml tube
inside 1.5 ml tube and pulse spin to rid of water. Discard outer tube and
replace.

4.Place agarose (preferably 1.0% HMP) in 0.6 ml tube and spin 20 mins at
6500 rpm. Collect eluate.

5.Can use eluate for PCR, ligations, etcc. directly. Alternatively you can
precipitate it to clean it.

Someone brought this protocol to the lab - where it comes from nobody
knows, if someone has the ref. please post it or e-mail it to me at:

leach at mbcrr.harvard.edu



Everybody must check out the newsgroup alt.netgames.bolo, download the game
bolo from sumex- etc.. and see you on the battlefield.... Mega smegging
fun.


Martin Leach

a.k.a Bulldozer



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