Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI18417
Trapped Gene
Aes (ENSMUSG00000054452)
Vector Insertion
Chr 10: 81028112 - 81028216
Public Clones not available
Private Clones OST423830 (lexicon)
Severity of mutation (?) Insertion after 64% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000102269 (Chr10:81028037..81028111 +)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
GCTCCTGAGCTGAACTCCAT Chr10:81028085..81028104 59.56 55 TGGCGAAAGGGGGATGTG Gene trap vector
 
 
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000102269 (Chr10:81028037..81028111 +)
Downstram Exon
ENSMUSE00000705336 (Chr10:81028217..81028438 +)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
GCTCCTGAGCTGAACTCCAT Chr10:81028085..81028104 59.56 55 AATCCGACTTCTCTCCGTCA Chr10:81028439..81028458 59.8 50

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000641506 Chr10:81022348..81022413 CGCGACTGACATGATGTTTC Chr10:81022377..81022396 60.27 50
upstream ENSMUSE00000665809 Chr10:81023923..81023964 No primer for this exon
upstream ENSMUSE00000298746 Chr10:81023964..81024061 CAAGTTCACCACCTCCGACT Chr10:81023990..81024009 60.15 55
upstream ENSMUSE00000665808 Chr10:81023967..81024061 CAAGTTCACCACCTCCGACT Chr10:81023990..81024009 60.15 55
upstream ENSMUSE00000574607 Chr10:81026840..81026903 GCCAGCGAGAAGTCAGAGAT Chr10:81026865..81026884 59.71 55
upstream ENSMUSE00000574606 Chr10:81027418..81027462 CCTACGGATTGAACATCGAGA Chr10:81027431..81027451 60.08 47.62
upstream ENSMUSE00000298726 Chr10:81027593..81027655 TGAAGAGGCTGAATGGGATT Chr10:81027603..81027622 59.63 45
upstream ENSMUSE00000102269 Chr10:81028037..81028111 GCTCCTGAGCTGAACTCCAT Chr10:81028085..81028104 59.56 55

*** Putative Vector Insertion (Chr 10: 81028112 - 81028216) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000368559 Chr10:81028217..81028909 CCTTCCCCAAGAGCTAATCC Chr10:81028454..81028473 60.03 55
downstream ENSMUSE00000705336 Chr10:81028217..81028438 AATCCGACTTCTCTCCGTCA Chr10:81028439..81028458 59.8 50

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GAGAGAGCCAAGCAGGTCAC Chr10:81028065..81028085 60.14 60 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GAGAGAGCCAAGCAGGTCAC Chr10:81028065..81028085 60.14 60 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000054452