Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI23797
Trapped Gene
Dppa2 (ENSMUSG00000072419)
Vector Insertion
Chr 16: 48314148 - 48314241
Public Clones (sanger)
Private Clones OST244565 (lexicon) OST199600 (lexicon) OST77509 (lexicon)
Severity of mutation (?) Insertion after 36% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000699688 (Chr16:48313996..48314147 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGCCTCCTATTAGGGACGTG Chr16:48314071..48314090 60.09 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000699688 (Chr16:48313996..48314147 +)
Downstram Exon
ENSMUSE00000699687 (Chr16:48314242..48314295 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGCCTCCTATTAGGGACGTG Chr16:48314071..48314090 60.09 55 GGAGTGTCTCCGAAGTCTCAA Chr16:48314279..48314299 59.44 52.38

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000699690 Chr16:48310590..48310652 TCATACTTCGGCCTGGAGAC Chr16:48310623..48310642 60.22 55
upstream ENSMUSE00000629029 Chr16:48311688..48311823 TTTAACGCTGGTCCCCTTTA Chr16:48311726..48311745 59.58 45
upstream ENSMUSE00000699688 Chr16:48313996..48314147 TGCCTCCTATTAGGGACGTG Chr16:48314071..48314090 60.09 55

*** Putative Vector Insertion (Chr 16: 48314148 - 48314241) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000699687 Chr16:48314242..48314295 GGAGTGTCTCCGAAGTCTCAA Chr16:48314279..48314299 59.44 52.38
downstream ENSMUSE00000699686 Chr16:48315775..48316030 GCTGCAAATGTGGACTCCTT Chr16:48315938..48315957 60.26 50
downstream ENSMUSE00000699684 Chr16:48317401..48317596 TACAGGCCGGTAACAGGAAG Chr16:48317544..48317563 60.12 55
downstream ENSMUSE00000699682 Chr16:48318746..48319625 GTTAAAATGCAACGGGCTGT Chr16:48319027..48319046 60 45

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 ATGGACACTAATCGCCTTGC Chr16:48314191..48314211 60.1 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GACACCGTGACTGGGAAAAC Chr16:48314194..48314214 60.41 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000072419