Stable Tandem Repeats?
Karthik Aghoram
kaghoram at unity.ncsu.edu
Wed Aug 22 08:29:15 EST 2001
Thanks for your informative post! The SURE cells worked for my inverted
repeat.
Regards,
Karthik
Robert Whittier (rfwhittier at hotmail.com) wrote:
: As pointed out by Duncan Clark in a similar vein in this
: month's thread on P1 lysogeny, you probably need to carry
: out trial and error tests with a variety of host strains.
: In my own limited experience, Stratagene's SURE cells were
: marginally more stable for inverted repeats, but Life
: Technologies' STABL2 were head-and-shoulders better for
: a construct carrying several separated direct repeats. Your
: mileage may vary. Both strains carry the gyrA96 mutation.
: Presumably this reduces supercoiling, although I have to
: admit ignorance whether this impacts the stability of direct
: repeats in any way. Supercoiling can be expected to promote
: chi structure formation in large palindromes. The centers
: of these should look just like Holliday structures, and as
: such are probably cleaved by the resolvases that normally
: mediate the final steps of genetic recombination. The two
: strains differ in that SURE attempts to knock down
: recombinantion with recB and recJ, while STABL2 uses recA1.
:
: Plasmid copy number and replicon types may also be major
: factors for stability, although they were not critical in
: my case.
:
: Assuming that you overcome the problem, please post what
: works back to this newsgroup.
:
: I have no affiliation with either of the companies mentioned
: above.
:
: /Bob
:
:
:
: _________________________________________________________________
: Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
:
: ---
--
-------------------------------------------------------------------------
Karthik Aghoram Postdoctoral Associate
Dept of Crop Science
e-mail: kaghoram at unity.ncsu.edu North Carolina St University
Phone (W): 919-515-2705 Raleigh, NC 27695-7620
-------------------------------------------------------------------------
More information about the Methods
mailing list