Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI25177
Trapped Gene
Arhgdia (ENSMUSG00000025132)
Vector Insertion
Chr 11: 120441059 - 120441280
Public Clones not available
Private Clones OST192835 (lexicon)
Severity of mutation (?) Insertion after 45% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000148095 (Chr11:120441281..120441364 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
TGTGACTCGACTGACCTTGG Chr11:120441323..120441342 59.86 55 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000148095 (Chr11:120441281..120441364 -)
Downstram Exon
ENSMUSE00000148094 (Chr11:120440982..120441058 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
TGTGACTCGACTGACCTTGG Chr11:120441323..120441342 59.86 55 ATCCGGTACTCCACACCTTC Chr11:120440976..120440995 58.87 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000645555 Chr11:120442836..120442916 GACGACGTTCGTCATTTGGT Chr11:120442872..120442891 60.97 50
upstream ENSMUSE00000668947 Chr11:120442304..120442694 GAATCTGCGGAACTGAAAGG Chr11:120442325..120442344 59.81 50
upstream ENSMUSE00000384199 Chr11:120441496..120441707 AAGAGCATCCAGGAGATCCA Chr11:120441570..120441589 59.76 50
upstream ENSMUSE00000717989 Chr11:120441496..120441707 AAGAGCATCCAGGAGATCCA Chr11:120441570..120441589 59.76 50
upstream ENSMUSE00000148095 Chr11:120441281..120441364 TGTGACTCGACTGACCTTGG Chr11:120441323..120441342 59.86 55

*** Putative Vector Insertion (Chr 11: 120441059 - 120441280) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000148094 Chr11:120440982..120441058 ATCCGGTACTCCACACCTTC Chr11:120440976..120440995 58.87 55
downstream ENSMUSE00000148096 Chr11:120440830..120440893 CCGGACACGATCTCTCTGTT Chr11:120440849..120440868 60.26 55
downstream ENSMUSE00000668948 Chr11:120439427..120440744 GAGATTCCACTCCCAGGACA Chr11:120440550..120440569 60.05 55
downstream ENSMUSE00000480495 Chr11:120439423..120440744 GAGATTCCACTCCCAGGACA Chr11:120440550..120440569 60.05 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 AGCTGGCATCTTGCCTCTTA Chr11:120441227..120441247 60.12 50 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 CATCTTGCCTCTCGTGACTG Chr11:120441221..120441241 59.57 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000025132