LINE inserts.

Joel Guss jg183 at columbia.edu
Fri Feb 6 23:25:51 EST 1998


Hi,
We are trying to find a PCR to genotype mutant mice with a LINE element
insert.  The primers to detect presense of a WT gene are on either
sides of where the insert is and work well.  One of the primers used
to detect the LINE insert is obviously in the LINE element.  PCR with
this primer, though, gives many nonspecific nonreproduceable bands
in both wild type and mutant mice!
This is most likely due to the presense of large numbers of these
inserts that contain very homologous sequences thorughout the genome.
Any primer we select even has at least three good anealing sites just
within the one LINE element we have!
Does anyone have any experience with this and might be able to lend
some advice?  We would prefer not to have to use Southerns to genotype
the mice.
Thanks.
                                                        Joel




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