Tools for the biologist enabling optimized use of gene trap clones

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UniTrap UNI24638
Trapped Gene
Mizf (ENSMUSG00000032119)
Vector Insertion
Chr 9: 44107876 - 44110492
Public Clones not available
Private Clones OST211899 (lexicon)
Severity of mutation (?) Insertion after 11% of polypeptide chain

Suggested RT-PCR (?)
Control PCR for the trapped allele
Upstream Exon
ENSMUSE00000372344 (Chr9:44110493..44110670 -)
Vector Sequence
Forward Primer Location Tm %GC LacZrt Reverse Primer Location
CACATGCATGGCTCTAAGGA Chr9:44110516..44110535 59.82 50 TGGCGAAAGGGGGATGTG Gene trap vector
Control PCR for the wild-type allele
Upstream Exon
ENSMUSE00000372344 (Chr9:44110493..44110670 -)
Downstram Exon
ENSMUSE00000263874 (Chr9:44107646..44107875 -)
Forward Primer Location Tm %GC Reverse Primer Location Tm %GC
CACATGCATGGCTCTAAGGA Chr9:44110516..44110535 59.82 50 GTAGACATGGCGGATGAGGT Chr9:44107774..44107793 59.96 55

Other possible Exon-Primers for RT-PCR (?)
  Exon ID Exon location Forward Primer Primer location Tm %GC
upstream ENSMUSE00000637489 Chr9:44113699..44113717 No primer for this exon
upstream ENSMUSE00000372344 Chr9:44110493..44110670 CACATGCATGGCTCTAAGGA Chr9:44110516..44110535 59.82 50

*** Putative Vector Insertion (Chr 9: 44107876 - 44110492) ***

  Exon ID Exon location Reverse Primer Primer location Tm %GC
downstream ENSMUSE00000263874 Chr9:44107646..44107875 GTAGACATGGCGGATGAGGT Chr9:44107774..44107793 59.96 55
downstream ENSMUSE00000263859 Chr9:44107296..44107407 GCCGATAGAACCACTCAGGA Chr9:44107352..44107371 60.22 55
downstream ENSMUSE00000263845 Chr9:44106956..44107108 GAAGCTTACACCGGTCCTTG Chr9:44107051..44107070 59.73 55
downstream ENSMUSE00000263818 Chr9:44106417..44106494 GCCTCTCTGTGGCAAATCTC Chr9:44106422..44106441 59.96 55
downstream ENSMUSE00000216701 Chr9:44106207..44106327 GTGATTCCGGAGGGAAGAAG Chr9:44106241..44106260 60.96 55
downstream ENSMUSE00000263776 Chr9:44105982..44106120 TCCGGAGGTCAATCAAATTC Chr9:44106073..44106092 59.87 45
downstream ENSMUSE00000263755 Chr9:44105776..44105900 CCACGTGTGAAGCATTTGTC Chr9:44105823..44105842 60.16 50
downstream ENSMUSE00000342535 Chr9:44102723..44104663 CTAAGCAGGCGAGAGCACTT Chr9:44103118..44103137 59.92 55

Suggested Genomic-PCR (?)
  Forward Forward location Tm %GC Reverse name Reverse Reverse Location
PCR 1 GGAGCTTAAGGGAGGTGTGA Chr9:44110457..44110477 59.28 55 L232 GATGTGCTGCAAGGCGATTA Gene trap vector
PCR 2 GCTACGTGACTGGGAAAACC Chr9:44110425..44110445 59.6 55 LacZ CCAGGGTTTTCCCAGTCACG Gene trap vector

Come back to gene ENSMUSG00000032119