TGGE vs DGGE &2D DGGE

WSchick at aol.com WSchick at aol.com
Thu Feb 29 20:15:06 EST 1996


In a message dated 96-02-29 14:39:25 EST, biometra at GATE.NET (Tony Sanchez)
writes:

>
>I am trying to decide between TGGE and DGGE for mutation screening. 

If you are screening for unknown mutations, you might also look into the 2D
DGGE method  commercialized by Ingeny BV of Leiden, The Netherlands.  FAX is
+31-71-21-02-36.  They have an automated instrument to do multiple sets of
both 1 & 2 D's, as well as single units.  

First diemension is regular electrophoresi, Second dimension is DGGE.
 Fragments (PCR or enzyme cut) are always in same horizontal size position.
 Mutations change the melting point and spots move up and down depending on
what has mutated.  They amplified all the exons in CF gene and discuovered a
few more mutations this way.  Less complex gels like this can be documented
by ethidium bromide. It is the onlyl theoretical 100% mutation method.  SSCP
hits only 60-70%.

For genomic samples, they also add an internal standard, probe it and use it
for adjusting other probe patterns.

Walt Schick




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