[bionet.molbio.methds-reagnts] Oligo Hyb Conditions

WOOLFORD.THOMPSON@BIONET-20.ARPA (06/02/88)

From: JOHN R. THOMPSON <WOOLFORD.THOMPSON@BIONET-20.ARPA>

I recall that a particular salt (the name of which escapes me) is 
advantageous to add to hybridizations involving oligomers.  It seems this 
salt minimizes GC bias and makes the Tm strictly dependent on length thereby 
decreasing false signals from sequences with one or two mismatches.  Can 
anyone give me a pointer to this?

Thanks, JRT

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BRUTLAG@BIONET-20.ARPA (06/05/88)

From: Doug Brutlag <brutlag@BIONET-20.ARPA>

John,

	I think that the salts that you are talking about for performing
hybridization are the tetraalkylamonium salts.  They are found to
decrease the difference in Tm between AT and GC bases and to allow very
specific hybridization of probes to complex genomic DNAs.

	Here are some references:

AN 88013408.  8801.
AU DiLella-A-G.  Woo-S-L.
TI Hybridization of genomic DNA to oligonucleotide probes in the
   presence of tetramethylammonium chloride.
SO Methods-Enzymol. 1987. 152. P 447-51.

AN 85166176.  8500.
AU Wood-W-I.  Gitschier-J.  Lasky-L-A.  Lawn-R-M.
IN Department of Molecular Biology, Genentech, Inc., South San
   Francisco, Ca 94080.
TI Base composition-independent hybridization in tetramethylammonium
   chloride: a method for oligonucleotide screening of highly complex
   gene libraries.
SO Proc-Natl-Acad-Sci-USA. 1985 Mar. 82(6). P 1585-8.

AN 81232835.  8100.
AU Wetmur-J-G.  Ruyechan-W-T.  Douthart-R-J.
IN Department of Microbiology, Mount Sinai School of Medicine of the
   City University of New York.
TI Denaturation and renaturation of Penicillium chrysogenum mycophage
   double-stranded ribonucleic acid in tetraalkylammonium salt
   solutions.
SO Biochemistry. 1981 May 26. 20(11). P 2999-3002.

AN 77222297.  7700.
AU Orosz-J-M.  Wetmur-J-G.
TI DNA melting temperatures and renaturation rates in concentrated
   alkylammonium salt solutions.
SO Biopolymers. 1977 Jun. 16(6). P 1183-99.

AN 76149669.  7600.
AU Voskoboinik-A-D.  Monaselidze-D-R.  Mgeladze-G-N.
   Chanchalashvili-Z-I.  Lazurkin-IuS.  Frank-kamenetskii-I-M.
TI [Study of DNA melting in the region of the inversion of relative
   stability of AT and GC pairs].
TT Issledovanie plavleniia DNK v oblasti inversii otnositel'noi
   stabil'nosti AT- i GC-PAR.
SO Mol-Biol (Mosk). 1975 Sep-Oct. 9(5). P 783-90.

	Hope this is what you were looking for.

Doug Brutlag
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