Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI30750
Trapped Gene
Plekhm1l (ENSMUSG00000051344)
Vector Insertion
Chr 1: 64968123 - 64985114
Public Clones IST14421H8 (tigm)
Private Clones not available
Severity of mutation (?) Insertion after 27% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000374303 (Chr1:64984271..64985113 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GGTGCTTTAAGAAGCGTTGC Chr1:64984693..64984712 60.03 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000374303 (Chr1:64984271..64985113 -)
Downstram Exon
ENSMUSE00000348305 (Chr1:64968124..64969056 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GGTGCTTTAAGAAGCGTTGC Chr1:64984693..64984712 60.03 50 TTTGGGGCTATCCAAGACAG Chr1:64968577..64968596 60.07 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000459706 Chr1:65003269..65003398 GAGAAGCCGCTTGTAAACAGA Chr1:65003311..65003331 59.65 47.62
upstream ENSMUSE00000374303 Chr1:64984271..64985113 GGTGCTTTAAGAAGCGTTGC Chr1:64984693..64984712 60.03 50
upstream ENSMUSE00000348305 Chr1:64968124..64969056 CTGTCTTGGATAGCCCCAAA Chr1:64968599..64968618 60.07 50

*** Putative Vector Insertion (Chr 1: 64968123 - 64985114) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000360192 Chr1:64939326..64939471 CCCGCTATAATTGCACACCT Chr1:64939391..64939410 59.98 50
downstream ENSMUSE00000345595 Chr1:64929703..64929896 GCTGGATGTCGATCAGAGGT Chr1:64929811..64929830 60.23 55
downstream ENSMUSE00000357386 Chr1:64907761..64907824 TGCAGGTCAGCGAGAGAGTA Chr1:64907743..64907762 59.88 55
downstream ENSMUSE00000395853 Chr1:64866510..64866667 TCCTCGAAGGGGTAGAGGAT Chr1:64866503..64866522 60.03 55
downstream ENSMUSE00000629817 Chr1:64835695..64841788 CTCCAAGATGCGTGCAGTAA Chr1:64840731..64840750 60.01 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 CAATGGACCTTGTGCTTCCT Chr1:64976063..64976083 60.11 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 TGATTACCCGTTCTGACGTG Chr1:64976059..64976079 59.57 50 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000051344