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
: 
: 
: 
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: ---

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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

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